Inositolphosphoryl ceramide (IPC) synthase genes from fungi

ABSTRACT

The invention provides isolated nucleic acid compounds encoding IPC synthase or subunit thereof from fungal cells. Also provided are vectors and transformed heterologous host cells for expressing IPC synthase and a method for identifying compounds that inhibit a fungal IPC synthase.

BACKGROUND OF THE INVENTION

[0001] This invention relates to recombinant DNA technology. Inparticular the invention pertains to the isolation of novel genes andproteins that encode IPC synthase or subunit thereof from a variety offungi and the use of said proteins in screens for inhibitors of IPCSynthase.

[0002] The incidence of life-threatening fungal infections is increasingat an alarming rate. With the exception of Staphylococci infections, thefungus C. albicans represents the fastest growing area of concern inhospital acquired bloodstream infections. About 90% of nosocomial fungalinfections are caused by species of Candida with the remaining 10% beingattributable to infections by Aspergillus, Cryptococcus, andPneumocystis. While effective antifungal compounds have been developedfor Candida there is growing concern that the rise in the incidence offungal infections may portend greater resistance and virulence in thefuture. Moreover, anti-Candida compounds frequently do not possessclinically significant activity against other fungal species.

[0003] Inositolphosphoryl ceramides are sphingolipids found in a numberof fungi including but not limited to S. cerevisiae, S. pombe, C.albicans, A. fumigatus, A. nidulans and H. capsulatum. A step ofsphingolipid biosynthesis unique to fungi and plants is catalyzed by theenzyme IPC synthase. The IPC synthase step, covalently links inositolphosphate and ceramide, and is essential for viability in S. cerevisiae.Although some elements of sphingolipid biosynthesis in fungi are sharedwith mammalian systems, the pathways diverge at the step after formationof ceramide. Thus, the formation of inositolphosphoryl ceramide isunique to fungi and plants, making IPC synthase a good molecular targetfor antifungal chemotherapy.

[0004] While compounds that target IPC synthase bode well for the futureof anti-fungal therapy, presently there are no clinically usefulcompounds that act at this step. Thus, there is a need for new compoundsthat inhibit IPC synthase.

BRIEF SUMMARY OF THE INVENTION

[0005] The present invention relates to fungal IPC synthase and toscreens for inhibitors thereof.

[0006] In one embodiment the invention relates to fungal genes thatencode IPC synthase, or subunit thereof.

[0007] In another embodiment, the invention relates to fungal IPCsynthase genes identified herein as SEQ ID NO:1, SEQ ID NO:4, SEQ IDNO:7, SEQ ID NO:10, SEQ ID NO:19, and SEQ ID NO:20.

[0008] In another embodiment, the invention relates to nucleic acidsthat are at least 70% homologous, and/or, will hybridize under highstringency conditions to a sequence identified herein as SEQ ID NO:1,SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:19, and SEQ ID NO:20.

[0009] In another embodiment the present invention pertains to theproteins produced by IPC synthase genes.

[0010] In yet another embodiment, the invention relates to proteinsdesignated herein as SEQ ID NO 2, SEQ ID NO 5, SEQ ID NO 8, SEQ ID NO11, and SEQ ID NO:21.

[0011] In still another embodiment the invention relates to the use ofpurified fungal IPC synthase or subunit thereof in high throughputscreens for inhibitors of fungal IPC synthase, said IPC synthase beingdesignated herein as SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ IDNO:11, SEQ ID NO:14, SEQ ID NO:17, and SEQ ID NO:21.

[0012] In another embodiment the invention relates to the use ofrecombinant host cells that carry a vector that expresses a fungal IPCsynthase in high throughput screens for inhibitors of fungal IPCsynthase.

[0013] In another embodiment the invention relates to the use of the IPCsynthase genes disclosed herein, or fragments thereof, as hybridizationprobes or PCR primers to identify and isolate homologous genes that arerelated in sequence and/or function.

DEFINITIONS

[0014] The terms “cleavage” or “restriction” of DNA refers to thecatalytic cleavage of the DNA with a restriction enzyme that acts onlyat certain sequences in the DNA (viz. sequence-specific endonucleases).The various restriction enzymes used herein are commercially availableand their reaction conditions, cofactors, and other requirements areused in the manner well known to one of ordinary skill in the art.Appropriate buffers and substrate amounts for particular restrictionenzymes are specified by the manufacturer or can readily be found in theliterature.

[0015] The term “fusion protein” denotes a hybrid protein molecule notfound in nature comprising a translational fusion or enzymatic fusion inwhich two or more different proteins or fragments thereof are covalentlylinked on a single polypeptide chain.

[0016] The term “plasmid” refers to an extrachromosomal genetic element.The starting plasmids herein are either commercially available, publiclyavailable on an unrestricted basis, or can be constructed from availableplasmids in accordance with published procedures. In addition,equivalent plasmids to those described are known in the art and will beapparent to the ordinarily skilled artisan.

[0017] “Recombinant DNA cloning vector” as used herein refers to anyautonomously replicating agent, including, but not limited to, plasmidsand phages, comprising a DNA molecule to which one or more additionalDNA segments can or have been added.

[0018] The term “recombinant DNA expression vector” as used hereinrefers to any recombinant DNA cloning vector, for example a plasmid orphage, in which a promoter and other regulatory elements are present toenable transcription of the inserted DNA.

[0019] The term “vector” as used herein refers to a nucleic acidcompound used for introducing exogenous DNA into host cells. A vectorcomprises a nucleotide sequence which may encode one or more proteinmolecules. Plasmid, cosmids, viruses, and bacteriophages, in the naturalstate or which have undergone recombinant engineering, are examples ofcommonly used vectors.

[0020] The terms “complementary” or “complementarity” as used hereinrefers to the capacity of purine and pyrimidine nucleotides to associatethrough hydrogen bonding in double stranded nucleic acid molecules. Thefollowing base pairs are complementary: guanine and cytosine; adenineand thymine; and adenine and uracil.

[0021] “Isolated nucleic acid compound” refers to any RNA or DNAsequence, however constructed or synthesized, which is locationallydistinct from its natural location.

[0022] A “primer” is a nucleic acid fragment which functions as aninitiating substrate for enzymatic or synthetic elongation of, forexample, a nucleic acid molecule.

[0023] The term “promoter” refers to a DNA sequence which directstranscription of DNA to RNA.

[0024] A “probe” as used herein is a nucleic acid compound thathybridizes with another nucleic acid compound, and is useful for blothybridizations, for example. A probe is at least 15 base pairs inlength, its sequence being at least 90% identical with the nucleic acidmolecules disclosed herein, or fragments thereof, or the complementsthereof. A probe may or may not be labeled with a detectable moiety. Asused herein, a probe is useful for hybridization analysis to identifysequences homologous to those disclosed herein.

[0025] The term “hybridization” as used herein refers to a process inwhich a single-stranded nucleic acid molecule joins with a complementarystrand through nucleotide base pairing. “Selective hybridization” refersto hybridization under conditions of high stringency. The degree ofhybridization depends upon, for example, the degree of complementarity,the stringency of hybridization, and the length of hybridizing strands.

[0026] The term “stringency” refers to hybridization conditions. Highstringency conditions disfavor non-homologous basepairing. Lowstringency conditions have the opposite effect. Stringency may bealtered, for example, by temperature and salt concentration.

[0027] “Low stringency” conditions comprise, for example, a temperatureof about 37° C. or less, a formamide concentration of less than about50%, and a moderate to low salt (SSC) concentration; or, alternatively,a temperature of about 50° C. or less, and a moderate to high salt(SSPE) concentration.

[0028] “High stringency” conditions comprise a temperature of about 42°C. or less, a formamide concentration of less than about 20%, and a lowsalt (SSC) concentration; or, alternatively, a temperature of about 65°C., or less, and a low salt (SSPE) concentration.

[0029] “SSC” comprises a hybridization and wash solution. 20×SSCcontains 3M sodium chloride, 0.3M sodium citrate, pH 7.0.

[0030] “SSPE” comprises a hybridization and wash solution. 1× SSPEcontains 180 mM NaCl, 9mM Na₂HPO₄, 0.9 mM NaH₂PO₄ and 1 mM EDTA, pH 7.4.

DETAILED DESCRIPTION OF THE INVENTION

[0031] In yeast and other fungi, for example C. albicans, Aspergillusnidulans, Aspergillus fumigatus, Cryptococcus neofomans, C. Krusei, C.parapsilosis, C. tropicalis, and C. glabrata, IPC synthase catalyzes astep in the synthesis of inositolphosphoryl ceramide from ceramide andphosphatidylinositol (G. Becker and R. Lester, Biosynthesis ofphosphoinositol-containing sphingolipid from phosphatidylinositol by amembrane prepartation from Saccharomyces cervisiae. J. Bacteriol. 142,747-754, 1980). Sphingolipids are necessary for growth and viability ofthe yeast S. cerevisiae. Since IPC synthase is unique to fungi andplants it is a good target for antifungal therapy in mammals.

[0032] The IPC synthase gene of C. glabrata comprises the DNA sequencedesignated herein as SEQ ID NO 1. The IPC synthase gene of C. Kruseicomprises the DNA sequence designated. herein as SEQ ID NO 4. The IPCsynthase gene of C. parapsilosis comprises the DNA sequence designatedherein as SEQ ID NO 7. The IPC synthase gene of C. tropicalis comprisesthe DNA sequence designated herein as SEQ ID NO 10. The IPC synthasegene of A. fumigatus is designated herein as SEQ ID NO:13. The IPCsynthase gene of A. nidulans is designated herein as SEQ ID NO:16. Thereare no intervening sequences in these genes. The IPC synthase gene of C.neoformans is designated herein as SEQ ID NO:19 and the cDNA thereof asSEQ ID NO:20. There is one intervening sequence in SEQ ID NO:19, frombase pair 1888 through base pair 1939. Those skilled in the art willrecognize that owing to the degeneracy of the genetic code (i.e. 64codons which encode 20 amino acids), numerous “silent” substitutions ofnucleotide base pairs could be introduced into these sequences withoutaltering the identity of the encoded amino acid(s) or protein products.All such substitutions are intended to be within the scope of theinvention. The genes disclosed and contemplated herein are useful forexpressing the protein encoded thereby, in vitro or in a recombinanthost cell.

[0033] Also contemplated is the use of said genes and fragments thereofas molecular hybridization probes for the identification and isolationof homologous genes.

[0034] Also contemplated by the present invention are nucleic acids thathybridize under high stringency conditions to the nucleic acid sequencesdisclosed herein.

[0035] Also contemplated are nucleic acids that are at least 70%identical in sequence to a nucleic acid sequence disclosed herein.

[0036] Gene Isolation Procedures

[0037] Those skilled in the art will recogize that the IPC synthasegenes disclosed herein may be obtained by a plurality of applicablegenetic and recombinant DNA techniques including, for example,polymerase chain reaction (PCR) amplification, or de novo DNA synthesis.(See e.g., J. Sambrook et al. Molecular Cloning, 2d Ed. Chap. 14(1989)).

[0038] Skilled artisans will recognize that the IPC synthase genesdisclosed herein, or fragments thereof, could be isolated by PCRamplification of genomic DNA isolated from suitable fungal cells usingoligonucleotide primers targeted to any suitable region of SEQ ID NO:1,SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, orSEQ ID NO:19. The skilled artisan understands that the choice ofsuitable primers may involve routine experimentation to achieve asuccessful outcome in a PCR amplification and that some trial and errorwith specific primers may be necessary. Methods for PCR amplificationare widely known in the art. See e.g. PCR Protocols: A Guide to Methodand Application, Ed. M. Innis et al., Academic Press (1990). The PCRamplification reaction comprises genomic DNA, suitable enzymes, primers,and buffers, and is conveniently carried out in a DNA Thermal Cycler(Perkin Elmer Cetus, Norwalk, Conn.). A positive result is determined bydetecting an appropriately-sized DNA fragment following agarose gelelectrophoresis.

[0039] Protein Production Methods

[0040] One of the embodiments of the present invention relates to thepurified proteins encoded by the IPC synthase genes disclosed herein, orfunctionally related proteins.

[0041] Skilled artisans will recognize that the proteins of the presentinvention can be synthesized by a variety of different methods. Forexample, the amino acid compounds of the invention can be made bychemical methods, well known in the art, including solid phase peptidesynthesis or recombinant methods. Both methods are described in U.S.Pat. No. 4,617,149, which hereby is incorporated by reference.

[0042] Solid phase chemical synthetic methods for polypeptides are wellknown in the art, and are described in general texts covering the area.See, e.g., H. Dugas and C. Penney, Bioorganic Chemistry (1981)Springer-Verlag, New York, 54-92. For example, peptides may besynthesized by solid-phase methodology utilizing an Applied Biosystems430A peptide synthesizer (Applied Biosystems, Foster City, Calif.) andsynthesis cycles supplied by Applied Biosystems. Protected amino acids,such as t-butoxycarbonyl-protected amino acids, and other reagents arecommercially available from many chemical supply houses.

[0043] The proteins of the present invention can also be produced byrecombinant DNA methods using a cloned IPC synthase gene describedherein. Recombinant methods are preferred if a high yield is desired.Expression of a cloned IPC synthase gene can be carried out in a varietyof suitable host cells, well known to those skilled in the art. For thispurpose, an IPC synthase gene is introduced into a host cell by anysuitable means, well known to those skilled in the art. Whilechromosomal integration of the cloned IPC synthase gene is within thescope of the present invention, it is preferred that the gene be clonedinto a suitable extra-chromosomally maintained expression vector so thatthe coding region of the IPC synthase gene is operably-linked to aconstitutive or inducible promoter.

[0044] The basic steps in the recombinant production of the IPC synthaseprotein are:

[0045] a) constructing a natural, synthetic or semi-synthetic DNAencoding IPC synthase;

[0046] b) incorporating said DNA into an expression vector in a mannersuitable for expressing an IPC synthase protein, either alone or as afusion protein;

[0047] c) transforming an appropriate eucaryotic or prokaryotic hostcell with said expression vector,

[0048] d) culturing said transformed host cell in a manner to expressthe IPC synthase protein; and

[0049] e) recovering membranes from said host cell and/or purifying theIPC synthase protein by any suitable means, well known to those skilledin the art.

[0050] Expressing Recombinant IPC synthase in Procaryotic and EucaryoticHost Cells

[0051] In general, prokaryotes are used for cloning DNA sequences andfor constructing the vectors of the present invention. Prokaryotes arealso employed in the production of the IPC synthase protein. Forexample, the Escherichia coli K12 strain 294 (ATCC No. 31446) isparticularly useful for expressing heterologous proteins in aprocaryotic host. Other strains of E. coli, bacilli such as Bacillussubtilis, enterobacteriaceae such as Salmonella typhimurium or Serratiamarcescans, various Pseudomonas species and other bacteria, such asStreptomyces, may also be employed as host cells in cloning andexpressing the recombinant proteins of this invention.

[0052] Promoters that are suitable for driving expression of genes inprokaryotes include b-lactamase [e.g. vector pGX2907, ATCC 39344,contains a replicon and b-lactamase gene], lactose systems [Chang etal., Nature_(London), 275:615 (1978); Goeddel et al., Nature (London),281:544 (1979)], alkaline phosphatase, and the tryptophan (trp) promotersystem [vector pATH1 (ATCC 37695) which is designed to facilitateexpression of an open reading frame as a trpE fusion protein under thecontrol of the trp promoter]. Hybrid promoters such as the tac promoter(isolatable from plasmid pDR540, ATCC-37282) are also suitable. Stillother bacterial promoters, whose nucleotide sequences are generallyknown, enable one of skill in the art to ligate such promoter sequencesto DNA encoding the proteins of the instant invention using linkers oradapters to supply any required restriction sites. Promoters for use inbacterial systems also will contain a Shine-Dalgarno sequenceoperably-linked to the DNA encoding the desired polypeptides. Theseexamples are illustrative rather than limiting.

[0053] The proteins of this invention may be synthesized either bydirect expression or as a fusion protein comprising the protein ofinterest as a translational fusion with another protein or peptide,which may be removable by enzymatic or chemical cleavage. It is oftenobserved in the production of certain peptides in recombinant systemsthat expression as a fusion protein prolongs the lifespan, increases theyield of the desired peptide, or provides a convenient means ofpurifying the protein. A variety of peptidases (e.g. enterokinase andthrombin) which cleave a polypeptide at specific sites or digest thepeptides from the amino or carboxy termini (e.g. diaminopeptidase) ofthe peptide chain are known. Furthermore, particular chemicals (e.g.cyanogen bromide) will cleave a polypeptide chain at specific sites. Theskilled artisan will appreciate the modifications necessary to the aminoacid sequence (and synthetic or semi-synthetic coding sequence ifrecombinant means are employed) to incorporate site-specific internalcleavage sites. See e.g., P. Carter, “Site Specific Proteolysis ofFusion Proteins”, Chapter 13, in Protein Purification: From MolecularMechanisms to Large Scale Processes, American Chemical Society,Washington, D.C. (1990).

[0054] In addition to prokaryotes, mammalian host cells and eucaryoticmicrobes, such as yeast, may also be used to express the proteins ofthis invention. The simple eucaryote Saccharomyces cerevisiae is themost commonly used eucaryotic microorganism, although a number of otheryeasts, such as Kluyveromyces lactis, are also suitable. For expressionin Saccharomyces, the plasmid YRp7 (ATCC-40053), for example, may beused. See, e.g., D. Stinchcomb et al., Nature, 282:39 (1979); J.Kingsman et al., Gene, 7:141 (1979); S. Tschemper et al., Gene, 10:157(1980). Plasmid YRp7 contains the TRP1 gene which provides a selectablemarker for use in a trp1 auxotrophic mutant.

[0055] Purification of Recombinantly-Produced IPC Synthase

[0056] An expression vector carrying a cloned IPC synthase gene or cDNAfrom any of the fungi disclosed herein (SEQ ID NO:1, SEQ ID NO:4, SEQ IDNO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, or SEQ IDNO:20) is transformed or transfected into a suitable host cell usingstandard methods. Cells which contain the vector are propagated underconditions suitable for expression of the IPC synthase protein. If anIPC synthase gene is under the control of an inducible promoter thengrowth conditions would incorporate the appropriate inducer. Therecombinantly-produced IPC synthase protein may be purified fromcellular extracts of transformed cells by any suitable means. In apreferred method for protein purification, an IPC synthase gene used intransforming a host cell is modified at the 5′ end to incorporateseveral histidine residues at the amino terminus of the encoded IPCsynthase protein. This “histidine tag” enables a single-step proteinpurification method referred to as “immobilized metal ion affinitychromatography” (IMAC), essentially as described in U.S. Pat. No.4,569,794, which hereby is incorporated by reference. The IMAC methodenables rapid isolation of substantially pure protein.

[0057] IPC synthase activity can be detected in membranes fromrecombinant cells transformed with the genes disclosed herein, or in themembranes of non-transformed fungal cells that express said genes. Saidmembranes are a useful source of IPC synthase activity and can be usedas a reagent in an assay for IPC synthase activity.

[0058] Other embodiments of the present invention relate to isolatednucleic acid sequences which encode SEQ ID NO:2, SEQ ID NO:5, SEQ IDNO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, or SEQ ID NO:21 orfragments thereof. As skilled artisans will recognize, the amino acidcompounds of the invention can be encoded by a multitude of differentnucleic acid sequences due to the degeneracy of the genetic code.Because these alternative nucleic acid sequences would encode the sameamino acid sequences, the present invention further comprises thesealternate nucleic acid sequences.

[0059] The IPC synthase genes comprising the present invention may beproduced using synthetic methods well known in the art. See, e.g., E. L.Brown, R. Belagaje, M. J. Ryan, and H. G. Khorana, Methods inEnzymology, 68:109-151 (1979). The DNA segments corresponding to an IPCsynthase gene could be generated using a conventional DNA synthesizingapparatus, such as the Applied Biosystems Model 380A or 380B DNAsynthesizers (Applied Biosystems, Inc., 850 Lincoln Center Drive, FosterCity, Calif. 94404) which employ phosphoramidite chemistry.Alternatively, phosphotriester chemistry may be employed to synthesizethe nucleic acids of this invention. [See, e.g., M. J. Gait, ed.,Oligonucleotide Synthesis, A Practical Approach, (1984).]

[0060] In an alternative method, IPC synthase DNA sequences comprising aportion or all of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10,SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, or SEQ ID NO:20 can begenerated from fungal genomic DNA using suitable oligonucleotide primerscomplementary to these sequences or region therein, utilizing thepolymerase chain reaction as described in U.S. Pat. No. 4,889,818, whichis incorporated herein by reference. Protocols for performing the PCRare disclosed in, PCR Protocols: A Guide to Method and Applications, Ed.Michael A. Innis et al., Academic Press, Inc. (1990), which hereby isincorporated by reference.

[0061] The ribonucleic acids of the present invention may be preparedusing polynucleotide synthetic methods discussed supra, or they may beprepared enzymatically using RNA polymerases to transcribe a DNAtemplate.

[0062] The most preferred system for preparing the ribonucleic acids ofthe present invention employs the RNA polymerase from the bacteriophageT7 or bacteriophage SP6. These RNA polymerases are highly specific andrequire the insertion of bacteriophage-specific sequences at the 5′ endof the template to be transcribed. See, J. Sambrook, et al., supra, at18.82-18.84.

[0063] This invention also provides nucleic acids, RNA or DNA, which arecomplementary to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQID NO:7, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:13, SEQ IDNO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, or SEQ IDNO:22.

[0064] The present invention also provides probes and primers useful formolecular biology techniques. A compound that is SEQ ID NO:1, SEQ IDNO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:12, SEQ ID NO:19, SEQ ID NO:20, or SEQ ID NO:22, or acomplementary sequence thereof, or a fragment thereof, and which is atleast 15 base pairs in length, and which will selectively hybridize toC. glabrata, C. krusei, C. parapsilosis, C. tropicalis or C. neoformansDNA or mRNA encoding IPC synthase, is provided. Preferably, the compoundis DNA.

[0065] The probes and primers contemplated herein can be preparedenzymatically by well known methods (See e.g. Sambrook et al. supra).

[0066] Another aspect of the present invention relates to recombinantDNA cloning vectors and expression vectors comprising the nucleic acidsdescribed and contemplated herein. Many of the vectors encompassedwithin this invention are described above. The preferred nucleic acidvectors are those which comprise DNA. The most preferred recombinant DNAvectors comprise the isolated DNA sequences SEQ ID NO:1, SEQ ID NO:4,SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, orSEQ ID NO:20.

[0067] The skilled artisan understands that choosing the mostappropriate cloning vector or expression vector depends upon a number offactors including the availability of appropriate restriction enzymesites, the type of host cell into which the vector is to be transfectedor transformed, the purpose of the transfection or transformation (e.g.,stable transformation as an extrachromosomal element, or integrationinto the host chromosome), the presence or absence of readily assayableor selectable markers (e.g., antibiotic resistance markers, metabolicmarkers, or the like), and the number of copies of the gene to bepresent in the host cell.

[0068] Vectors suitable to carry the nucleic acids of the presentinvention include RNA viruses, DNA viruses, lytic bacteriophages,lysogenic bacteriophages, stable bacteriophages, plasmids, viroids, andthe like. The most preferred vectors are plasmids.

[0069] When preparing an expression vector the skilled artisanunderstands that there are many variables to be considered, for example,whether to use a constitutive or inducible promoter. Inducible promotersare preferred because they may be the basis for high level andregulatable expression of an operably-linked gene. The skilled artisanwill recognize a number of inducible promoters and inducers, forexample, carbon source, metal ions, heat, and others known to theskilled artisan. The practitioner also understands that the amount ofnucleic acid or protein to be produced dictates, in part, the selectionof the expression system. The addition of certain nucleotide sequences,such as a sequence encoding a signal peptide preceding the codingsequence, is useful to direct localization of the resulting polypeptide.

[0070] Host cells harboring the nucleic acids disclosed herein are alsoprovided by the present invention. A preferred host is E. coli, intowhich has been transfected or transformed a vector that comprises anucleic acid of the present invention.

[0071] The present invention also provides a method for constructing arecombinant host cell capable of expressing SEQ ID NO:2, SEQ ID NO:5,SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, or SEQ ID NO:21said method comprising transforming a host cell with a recombinant DNAvector that comprises an isolated DNA sequence that encodes one of thesesequences. Suitable host cells include any strain of E. coli or fungalcell that can accomodate high level expression of a gene(s) introducedby transformation or transfection. Preferred vectors for expression arethose that comprise SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10,SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, or SEQ ID NO:20. Transformedhost cells may be cultured under conditions well known to skilledartisans such that SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11,SEQ ID NC):14, SEQ ID NO:17, or SEQ ID NO:21 is expressed, therebyproducing fungal IPC synthase protein in a recombinant host cell.

[0072] For the purpose of identifying or developing antifungalcompounds, it would be desirable to determine those agents which inhibitIPC synthase activity. A method for determining whether a substance willinhibit the enzymatic reaction catalyzed by IPC synthase comprisescontacting a source of IPC synthase activity (e.g. membrane preparationfrom a cell that expresses IPC synthase), or a purified IPC synthaseprotein, or fragment exhibiting said synthase activity, with a testsubstance and monitoring IPC synthase activity by any suitable means.

[0073] The instant invention provides such a screening system useful fordiscovering agents which inhibit an IPC synthase, said screening systemcomprising the steps of:

[0074] a) preparing a source of IPC synthase, or IPC synthase protein,or subunit thereof;

[0075] b) exposing said IPC synthase or source thereof to a testinhibitor;

[0076] c) introducing substrate; and

[0077] d) quantifying the loss of activity of said IPC synthase.

[0078] Utilization of the screening system described above provides ameans to determine compounds that may interfere with fungal sphingolipidbiosynthesis. This screening system may be adapted to automatedprocedures such as a PANDEX® (Baxter-Dade Diagnostics) system allowingfor efficient high-volume screening of potential therapeutic agents.

[0079] In such a screening protocol IPC synthase or subunit thereof isprepared as described herein, preferably using recombinant DNAtechnology. Alternatively, the reaction can be carried out usingmembranes from cells that express ICP synthase, as a source of IPCsynthase activity. Preferably, the cells are recombinant cells thatincorporate a recombinantly-expressed IPC synthase into the cellmembrane. Most preferably, the recombinant cells are yeast cells. Asample of a test compound is then introduced into a reaction vesselcontaining IPC synthase activity, followed by the addition of enzymesubstrate. Alternatively, substrate may be added simultaneously with thetest compound.

[0080] The following examples more fully describe the present invention.Those skilled in the art will recognize that the particular reagents,equipment, and procedures described are merely illustrative and are notintended to limit the present invention in any manner.

EXAMPLE 1 Construction of a DNA Vector for Expressing the C. glabrataIPC synthase Gene in Homologous or Heterologous Host

[0081] An expression vector suitable for expressing the IPC synthasegene of C. glabrata (SEQ ID NO:1) in E. coli contains an origin ofreplication (Ori), an ampicillin resistance gene (Amp) useful forselecting cells that have incorporated the vector followingtranformation. The vector also includes the T7 promoter and T7terminator sequences in operable linkage to the coding region of the IPCsynthase gene. Parent plasmid pET11A (obtained from Novogen, Madison,Wis.) is linearized by digestion with appropriate endonucleases andligated to a DNA fragment comprising the coding region of the C.glabrata IPC synthase gene.

[0082] The IPC synthase gene ligated into the expression vector ismodified at the 5′ end (amino terminus of encoded protein) in order tosimplify purification of the encoded IPC synthase protein product. Forthis purpose, an oligonucleotide encoding 8 histidine residues and afactor Xa cleavage site is inserted after the ATG start codon atnucleotide positions 1 to 3 of SEQ ID NO:1. Placement of the histidineresidues at the amino terminus of the encoded protein enables the IMACone-step protein purification procedure (See below).

EXAMPLE 2 Expression of C. glabrata IPC synthase Gene in Echerichia coliand Purification of IPC Synthase Enzyme

[0083] A plasmid from Example 1 is transformed into E. coli BL21(DE3)(hsdS gal lcIts857 ind1Sam7nin5lacUVS-T7gene 1) using standardmethods (See e.g. Sambrook et al. Supra). Transformants, selected forresistance to ampicillin, are chosen at random and tested for thepresence of the vector by agarose gel electrophoresis using quickplasmid preparations. Colonies that contain the vector are grown,processed, and the protein encoded by the IPC synthase gene is purifiedby immobilized metal ion affinity chromatography (IMAC), essentially asdescribed in U.S. Pat. No. 4,569,794, the entire contents of which ishereby incorporated by reference.

[0084] Briefly, the IMAC column is prepared as follows. A metal-freechelating resin (e.g. SEPHAROSE 6B IDA, Pharmacia) is washed indistilled water to remove preservative substances and infused with asuitable metal ion [e.g. Ni(II), Co(II), or Cu(II)] by adding a 50 mMmetal chloride or metal sulfate aqueous solution until about 75% of theinterstitial spaces of the resin is saturated with colored metal ion.The column is then ready to receive a crude cellular extract containingthe IPC synthase protein product encoded by the vector.

[0085] After removing unbound proteins and other materials by washingthe column with suitable buffer, pH 7.5, the bound protein is eluted inbuffer at pH 4.3 essentially as described in U.S. Pat. No. 4,569,794.

EXAMPLE 3 Biochemical Assay for Inhibitors of IPC Synthase Using FungalMembrane Preparations

[0086] The activity of the IPC synthase enzyme is assayed by preparingmembranes from C. glabrata, for example, and using said membranes as asource of IPC synthase activity.

[0087] A suitable, rich medium, for example YEPD, is innoculated with aculture and allowed to grow overnight at room temperature with vigorousshaking. About 250 ml of fresh medium containing 20 ug/ml myo-inositolis innoculated with the overnight culture, and grown overnight at 30° C.Cells are harvested by centrifugation and resuspended in ice cold 50 mMpotassium phosphate buffer, pH 7. Cells are washed twice in the samebuffer and then resuspended in the same buffer containing 5 mMdithiothreitol (DTT), 1 ug,/ml aprotinin, 0.6 uM leupeptin, 1 mM PMSF,and 1 ug/ml pepstatin A. Cells are ruptured using glass beads in aprocedure that involves 5 successive vortexings each for 30 secondsfollowed by 2 to 5 minute intervals of rest on ice. Membranes arepelleted by centrifugation at 100,000×g for 1 hour at 4° C. The pelletis resuspended in cold buffer containing DTT and protease inhibitors anddisrupted further by Dounce homogenization with 5 to 6 strokes on ice.Membranes are mixed with glycerol to a final concentration of 33% andstored frozen at −80° C. For use as “protein stock,” thaw and dilute to4.8 mg/ml in 0.05 M potassium phosphate (Kpi) buffer, pH 7.0 on ice.

[0088] IPC synthase is assayed by any suitable method. For example:

[0089] Final Assay Conditions:

[0090] 5-50 uM NBD-C6-ceramide (from Molecular Probes)

[0091] 1 mM phosphatidylinositol (Sigma-soybean)

[0092] 2 mM CHAPS

[0093] 50 mM potassium phosphate buffer, pH 7.0

[0094] Up to 2.5% organic co-solvent

[0095] Protein to 1.2 mg/ml

[0096] Final Vol. 100 ul

[0097] 2X Master Mix (for 15 samples)

[0098] 46 ul of 1 mg/ml NBD-C6-ceramide in MeOH

[0099] 142 ul of 10 mg/ml phosphatidylinositol in CHCl₃

[0100] Dry under vacuum

[0101] Redissolve by adding in order

[0102] 160 ul 20 mM CHAPS

[0103] 560 ul deionized water

[0104] 80 ul 0.5 M Kpi buffer, pH 7.0

[0105] Sonicate 5 minutes.

[0106] Aliquot 50 ul of 2X master mix to screw cap microfuge at roomtemperature. Add 20 ul of 0.05 M Kpi buffer pH 7.0 to each samplefollowed by 5 ul of test compound or extract. The reaction is initiatedby adding 25 ul of protein stock and incubating for 1 hour in a 30° C.water bath. Reactions are terminated by adding 900 ul of cold absolutemethanol. Samples are stored at −20° C. for 1 hour and centrifuged at14000×g at 4° C. for 30 minutes.

[0107] The reaction is monitored by HPLC on a Beckman Ultrasphere-ODS 5um, 150×4 mm column. The chromatogram is developed in 87% MeOH/13% 50 mMTEAP, pH 5.8, at a flow rate of 1 ml/min at room temperature through aBeckman Model 157 Fluorescence detector, usingNBD-C₆-phosphatidylcholine as an internal standard.

[0108] Inhibition studies are carried out using the same reactionconditions described in the preceding paragraph. Compounds to be studiedfor inhibitory activity are added to final concentrations of between 1pM and 10 mM.

EXAMPLE 4 Expression of C. parapsilosis (SEQ ID NO:7) IPC Synthase Genein S. cerevisiae

[0109] A yeast/E. coli shuttle vector suitable for expressing the C.parapsilosis IPC synthase gene (SEQ ID NO:7) in S. cerevisiae isconstructed in parent plasmid YEp351 (See J. Hill et. al., Yeast, 2,163-167, 1986), which contains the multiple cloning region of pUC18, theAmp^(R) gene for selection in E. coli, 2μ replicon, and the LEU2 genefor selection in yeast. A fragment containing the C. parapsilosis IPCsynthase gene is prepared by PCR amplification. Suitable primers to the5′ and 3′ ends of the coding region disclosed in SEQ ID NO:7 areconstructed to contain BamH1 cloning sites in addition to IPC synthasecoding information. After PCR amplification of C. parapsilosis genomicDNA, the amplified fragment is purified by any sutiable method, forexample, gel purification of an appropriately sized fragment, followedby treatment with restriction enzyme BamH1, and ligation to BamH1digested YEp351. The recombinant plasmid carrying the IPC synthase geneis transformed into any suitable leu strain of S. cerevisiae andtransformants selected for growth in a medium that lacks added leucine.Membranes from tranformants are prepared, and IPC synthase activityassayed as in Example 3.

1 24 1 2000 DNA Candida glabrata CDS (501)..(1748) 1 aaaacgtcattttaaaatta ggctttagat tgtaactcag atgattttcg tcaatccagt 60 gtacgaaacttggcaaatgc taaaacttca acaaataaat catcatccct aatactagtg 120 taaataatttaatgtacatg gtgttctatt tctaatgtag aaatatgtat atgacattag 180 atcaactcatattgttcaaa actctgtcaa aataatttag tgaggcaaag gtgtcgtccc 240 ccgaatgaacttgtaactca ccttgaagga gatatttgca tttacttgat aatccgaaac 300 ttttgcaacttaacatttta aaactagaag ttatttggta tctggtatta atttttatta 360 gctttgctgattacgacacg ttacgaagga agatattgac gctcagaatt tttatttagg 420 agaaatttatcataatagac aatatttgcc attaccgatt tctaactaaa tatcataatc 480 atagttattagacaaattca atg aga ctt gga aga tgg ttt gct att ttg agc 533 Met Arg LeuGly Arg Trp Phe Ala Ile Leu Ser 1 5 10 tca aaa tta tgc tca gat aca gatgta act aag ttg gag aga agt ctt 581 Ser Lys Leu Cys Ser Asp Thr Asp ValThr Lys Leu Glu Arg Ser Leu 15 20 25 gat gta agg gtc aca tta aga cga ctcagg agt tac aga ccc aca ata 629 Asp Val Arg Val Thr Leu Arg Arg Leu ArgSer Tyr Arg Pro Thr Ile 30 35 40 ggt gac aca gtt cat tat tca ttt ttg tcaatg att cta ttg tat tca 677 Gly Asp Thr Val His Tyr Ser Phe Leu Ser MetIle Leu Leu Tyr Ser 45 50 55 tat ttt gct aat cca cta cca ttc att gtg aaaaca ttg ata gcg atg 725 Tyr Phe Ala Asn Pro Leu Pro Phe Ile Val Lys ThrLeu Ile Ala Met 60 65 70 75 att tta atg act ctg ttt gtc gtc ccg atg acatct caa ttt ttc ttt 773 Ile Leu Met Thr Leu Phe Val Val Pro Met Thr SerGln Phe Phe Phe 80 85 90 aat gca ctt cca ata ttg aca tgg cta atc ttg tatttt aca tca tca 821 Asn Ala Leu Pro Ile Leu Thr Trp Leu Ile Leu Tyr PheThr Ser Ser 95 100 105 cat att cct aat agt cat aga cct tca att tct gttgag gtg ctt cct 869 His Ile Pro Asn Ser His Arg Pro Ser Ile Ser Val GluVal Leu Pro 110 115 120 gcc att gaa act atc ctg tat ggc gat aac tta agtgag att ttg gct 917 Ala Ile Glu Thr Ile Leu Tyr Gly Asp Asn Leu Ser GluIle Leu Ala 125 130 135 gca tgg caa aat aca att cta gat atc cta gct tggatt cca tat ggt 965 Ala Trp Gln Asn Thr Ile Leu Asp Ile Leu Ala Trp IlePro Tyr Gly 140 145 150 155 ctg ttt cac ttt ggc gca cca ttt gtg gtt gcaatt gtt tta ttt ata 1013 Leu Phe His Phe Gly Ala Pro Phe Val Val Ala IleVal Leu Phe Ile 160 165 170 ttt gga cca ccg aca gtt ctt cag ggt tat gcgttt gcg ttt ggt tat 1061 Phe Gly Pro Pro Thr Val Leu Gln Gly Tyr Ala PheAla Phe Gly Tyr 175 180 185 gtg aat tta ttt ggg gtg tta ttc caa aac atattt ccc gcc gcg gct 1109 Val Asn Leu Phe Gly Val Leu Phe Gln Asn Ile PhePro Ala Ala Ala 190 195 200 cct tgg tat aaa ata cat tat gga ctc caa gttgca aat tac agt atc 1157 Pro Trp Tyr Lys Ile His Tyr Gly Leu Gln Val AlaAsn Tyr Ser Ile 205 210 215 aaa ggt atg cct ggt gga ctt tct aga att gatgat ctt ttg ggt gtg 1205 Lys Gly Met Pro Gly Gly Leu Ser Arg Ile Asp AspLeu Leu Gly Val 220 225 230 235 aat tta tat tca agt ggt ttt aag aac tctgct gtc gtt ttt gga gct 1253 Asn Leu Tyr Ser Ser Gly Phe Lys Asn Ser AlaVal Val Phe Gly Ala 240 245 250 ttt cct tct ctt cat tct gca tgt gca actatg gaa gca ctc ttc ttc 1301 Phe Pro Ser Leu His Ser Ala Cys Ala Thr MetGlu Ala Leu Phe Phe 255 260 265 tca tat tgt ttt cca agc ttg aca ccg ctcttt att ttc tac gtt tgg 1349 Ser Tyr Cys Phe Pro Ser Leu Thr Pro Leu PheIle Phe Tyr Val Trp 270 275 280 tgg tta tgg tgg tct aca atg tat ctt actcat cat tat ttt gtt gat 1397 Trp Leu Trp Trp Ser Thr Met Tyr Leu Thr HisHis Tyr Phe Val Asp 285 290 295 cta aca gca ggt tct gtt ttg gct tat ttcata ttt caa tat acc aaa 1445 Leu Thr Ala Gly Ser Val Leu Ala Tyr Phe IlePhe Gln Tyr Thr Lys 300 305 310 315 tac aca cat cta cct att gtt gat ccaaat ctg ctc ttc cgt tgg tcc 1493 Tyr Thr His Leu Pro Ile Val Asp Pro AsnLeu Leu Phe Arg Trp Ser 320 325 330 tac gac agt att gaa ttt tac gat gttaag aac aat gac cct tta tcc 1541 Tyr Asp Ser Ile Glu Phe Tyr Asp Val LysAsn Asn Asp Pro Leu Ser 335 340 345 att aca aat aac gat ata gag agt atacct cta aac acc att gaa cta 1589 Ile Thr Asn Asn Asp Ile Glu Ser Ile ProLeu Asn Thr Ile Glu Leu 350 355 360 gat aca gaa gaa gtg ttc aat att gagcgg gaa aaa tct agg aca cca 1637 Asp Thr Glu Glu Val Phe Asn Ile Glu ArgGlu Lys Ser Arg Thr Pro 365 370 375 gaa ttg tca cat gca gtg tct tca tcttct cta gct aac ggt cgc tct 1685 Glu Leu Ser His Ala Val Ser Ser Ser SerLeu Ala Asn Gly Arg Ser 380 385 390 395 caa aca tat cca aat act gga aataca ttc aaa tcc aag aca aga ctg 1733 Gln Thr Tyr Pro Asn Thr Gly Asn ThrPhe Lys Ser Lys Thr Arg Leu 400 405 410 agt aaa ctg gtc att taaaaccagtatttataaat agagactatc ttcttctact 1788 Ser Lys Leu Val Ile 415 gtacatacaatgatctttct atattctaac gaaatcaata agtgcatact atattcattt 1848 aacatgatcaatattattta acagatccat taggtgctgg cttcccaaga tagttaccaa 1908 aacgataattttttctcaaa atttgccctt tgagaaaaac ctccgtctag tcacccctta 1968 ccacacagcacaaactgaaa atcgatggga ta 2000 2 416 PRT Candida glabrata 2 Met Arg LeuGly Arg Trp Phe Ala Ile Leu Ser Ser Lys Leu Cys Ser 1 5 10 15 Asp ThrAsp Val Thr Lys Leu Glu Arg Ser Leu Asp Val Arg Val Thr 20 25 30 Leu ArgArg Leu Arg Ser Tyr Arg Pro Thr Ile Gly Asp Thr Val His 35 40 45 Tyr SerPhe Leu Ser Met Ile Leu Leu Tyr Ser Tyr Phe Ala Asn Pro 50 55 60 Leu ProPhe Ile Val Lys Thr Leu Ile Ala Met Ile Leu Met Thr Leu 65 70 75 80 PheVal Val Pro Met Thr Ser Gln Phe Phe Phe Asn Ala Leu Pro Ile 85 90 95 LeuThr Trp Leu Ile Leu Tyr Phe Thr Ser Ser His Ile Pro Asn Ser 100 105 110His Arg Pro Ser Ile Ser Val Glu Val Leu Pro Ala Ile Glu Thr Ile 115 120125 Leu Tyr Gly Asp Asn Leu Ser Glu Ile Leu Ala Ala Trp Gln Asn Thr 130135 140 Ile Leu Asp Ile Leu Ala Trp Ile Pro Tyr Gly Leu Phe His Phe Gly145 150 155 160 Ala Pro Phe Val Val Ala Ile Val Leu Phe Ile Phe Gly ProPro Thr 165 170 175 Val Leu Gln Gly Tyr Ala Phe Ala Phe Gly Tyr Val AsnLeu Phe Gly 180 185 190 Val Leu Phe Gln Asn Ile Phe Pro Ala Ala Ala ProTrp Tyr Lys Ile 195 200 205 His Tyr Gly Leu Gln Val Ala Asn Tyr Ser IleLys Gly Met Pro Gly 210 215 220 Gly Leu Ser Arg Ile Asp Asp Leu Leu GlyVal Asn Leu Tyr Ser Ser 225 230 235 240 Gly Phe Lys Asn Ser Ala Val ValPhe Gly Ala Phe Pro Ser Leu His 245 250 255 Ser Ala Cys Ala Thr Met GluAla Leu Phe Phe Ser Tyr Cys Phe Pro 260 265 270 Ser Leu Thr Pro Leu PheIle Phe Tyr Val Trp Trp Leu Trp Trp Ser 275 280 285 Thr Met Tyr Leu ThrHis His Tyr Phe Val Asp Leu Thr Ala Gly Ser 290 295 300 Val Leu Ala TyrPhe Ile Phe Gln Tyr Thr Lys Tyr Thr His Leu Pro 305 310 315 320 Ile ValAsp Pro Asn Leu Leu Phe Arg Trp Ser Tyr Asp Ser Ile Glu 325 330 335 PheTyr Asp Val Lys Asn Asn Asp Pro Leu Ser Ile Thr Asn Asn Asp 340 345 350Ile Glu Ser Ile Pro Leu Asn Thr Ile Glu Leu Asp Thr Glu Glu Val 355 360365 Phe Asn Ile Glu Arg Glu Lys Ser Arg Thr Pro Glu Leu Ser His Ala 370375 380 Val Ser Ser Ser Ser Leu Ala Asn Gly Arg Ser Gln Thr Tyr Pro Asn385 390 395 400 Thr Gly Asn Thr Phe Lys Ser Lys Thr Arg Leu Ser Lys LeuVal Ile 405 410 415 3 1248 RNA Candida glabrata 3 augagacuug gaagaugguuugcuauuuug agcucaaaau uaugcucaga uacagaugua 60 acuaaguugg agagaagucuugauguaagg gucacauuaa gacgacucag gaguuacaga 120 cccacaauag gugacacaguucauuauuca uuuuugucaa ugauucuauu guauucauau 180 uuugcuaauc cacuaccauucauugugaaa acauugauag cgaugauuuu aaugacucug 240 uuugucgucc cgaugacaucucaauuuuuc uuuaaugcac uuccaauauu gacauggcua 300 aucuuguauu uuacaucaucacauauuccu aauagucaua gaccuucaau uucuguugag 360 gugcuuccug ccauugaaacuauccuguau ggcgauaacu uaagugagau uuuggcugca 420 uggcaaaaua caauucuagauauccuagcu uggauuccau auggucuguu ucacuuuggc 480 gcaccauuug ugguugcaauuguuuuauuu auauuuggac caccgacagu ucuucagggu 540 uaugcguuug cguuugguuaugugaauuua uuuggggugu uauuccaaaa cauauuuccc 600 gccgcggcuc cuugguauaaaauacauuau ggacuccaag uugcaaauua caguaucaaa 660 gguaugccug guggacuuucuagaauugau gaucuuuugg gugugaauuu auauucaagu 720 gguuuuaaga acucugcugucguuuuugga gcuuuuccuu cucuucauuc ugcaugugca 780 acuauggaag cacucuucuucucauauugu uuuccaagcu ugacaccgcu cuuuauuuuc 840 uacguuuggu gguuaugguggucuacaaug uaucuuacuc aucauuauuu uguugaucua 900 acagcagguu cuguuuuggcuuauuucaua uuucaauaua ccaaauacac acaucuaccu 960 auuguugauc caaaucugcucuuccguugg uccuacgaca guauugaauu uuacgauguu 1020 aagaacaaug acccuuuauccauuacaaau aacgauauag agaguauacc ucuaaacacc 1080 auugaacuag auacagaagaaguguucaau auugagcggg aaaaaucuag gacaccagaa 1140 uugucacaug cagugucuucaucuucucua gcuaacgguc gcucucaaac auauccaaau 1200 acuggaaaua cauucaaauccaagacaaga cugaguaaac uggucauu 1248 4 2000 DNA Candida kruisii CDS(300)..(1739) 4 acagtaaaaa gaaattgcgc tgcatgaaac tttttttttt ttttttttcggtgtttcttc 60 tcgtttttct cgtttttctc gtttttctcg gtttcttttg agaatgtaatgaaatttgga 120 aactgtacaa acattctcag tgttgatatt aacaataata accacattgacattgacaac 180 atcattatta ggttctttcc atttgtatta tttccatctc ggacacgtcagctatagacc 240 gtcgacactc cctattccct gcaccaacta ggaagaacag gaagaacaagaagaacaga 299 atg ctc tcg ctc ccg gcc ctt att agg cgc ctg tat tcc tccttt att 347 Met Leu Ser Leu Pro Ala Leu Ile Arg Arg Leu Tyr Ser Ser PheIle 1 5 10 15 tcg gca aga gaa aag ggt aat tca ttg cct gca tta aag acatcc ttt 395 Ser Ala Arg Glu Lys Gly Asn Ser Leu Pro Ala Leu Lys Thr SerPhe 20 25 30 aac ccc ctc cct aac ttc aag acc ttc ccg gag aga ctg gtc tcggca 443 Asn Pro Leu Pro Asn Phe Lys Thr Phe Pro Glu Arg Leu Val Ser Ala35 40 45 tcc ata tcc aat gtg gtc cat ttc acg att atc ttt ctt gtc ttt gtc491 Ser Ile Ser Asn Val Val His Phe Thr Ile Ile Phe Leu Val Phe Val 5055 60 ttc tcc atc tca ata atg cct gta ccc cta ctt gtg aaa gcc att ctt539 Phe Ser Ile Ser Ile Met Pro Val Pro Leu Leu Val Lys Ala Ile Leu 6570 75 80 atc att gtc ttg att tac atc aac ttt gct cca att ttc gcc cag ttc587 Ile Ile Val Leu Ile Tyr Ile Asn Phe Ala Pro Ile Phe Ala Gln Phe 8590 95 ttt aac ggc ggc ttg ccg atc ctc tca tgg gtc att ttg ttc ttt aac635 Phe Asn Gly Gly Leu Pro Ile Leu Ser Trp Val Ile Leu Phe Phe Asn 100105 110 tca aag aac ata cct ctg tca atg aag aga ccc atc tcc gtc aag gtc683 Ser Lys Asn Ile Pro Leu Ser Met Lys Arg Pro Ile Ser Val Lys Val 115120 125 ttg ccc gca ttg gag aca atc ttc tat ggc gat aat tta tct gaa atc731 Leu Pro Ala Leu Glu Thr Ile Phe Tyr Gly Asp Asn Leu Ser Glu Ile 130135 140 ttg gct tct cat aca aac act ttc ttg gac ttg ttt gct tgg ttc cca779 Leu Ala Ser His Thr Asn Thr Phe Leu Asp Leu Phe Ala Trp Phe Pro 145150 155 160 tat ggt ata gtc cat ttc tcc ctg ccc ttt gtt gtg gca gcc cttatt 827 Tyr Gly Ile Val His Phe Ser Leu Pro Phe Val Val Ala Ala Leu Ile165 170 175 ttt tta ttt gcc ccc cct aaa acc tta ccc caa ttt tgt tgg tccttt 875 Phe Leu Phe Ala Pro Pro Lys Thr Leu Pro Gln Phe Cys Trp Ser Phe180 185 190 ggt tat atg aat ttg gtt ggc gtc att atc caa aac ttg gtt ttctca 923 Gly Tyr Met Asn Leu Val Gly Val Ile Ile Gln Asn Leu Val Phe Ser195 200 205 tgt gct cca cca tgg tat aaa gtg ctg cat ggg ttg gac aag gcaaac 971 Cys Ala Pro Pro Trp Tyr Lys Val Leu His Gly Leu Asp Lys Ala Asn210 215 220 tat tca atg aag gga tcg cca gga gga ttg ggt aga atc gat ggactg 1019 Tyr Ser Met Lys Gly Ser Pro Gly Gly Leu Gly Arg Ile Asp Gly Leu225 230 235 240 ttt ggt ttc gat atg tac acc tcg ggt ttc acc aat tct cccttg att 1067 Phe Gly Phe Asp Met Tyr Thr Ser Gly Phe Thr Asn Ser Pro LeuIle 245 250 255 ttc ggt gca atg ccc tcc ttg cat tcg gga tgt tca act atggac gca 1115 Phe Gly Ala Met Pro Ser Leu His Ser Gly Cys Ser Thr Met AspAla 260 265 270 tta tgg ttg tct tat cta ttc cct aaa ttc acc cct cta tttgtt ttc 1163 Leu Trp Leu Ser Tyr Leu Phe Pro Lys Phe Thr Pro Leu Phe ValPhe 275 280 285 tac gtt tgc tgg tta tgg ttc tct aca atg tat ttg act catcat tac 1211 Tyr Val Cys Trp Leu Trp Phe Ser Thr Met Tyr Leu Thr His HisTyr 290 295 300 ttc atc gat gtt gtt gtt ggt tct tgt ttg gca att gga ttcttc act 1259 Phe Ile Asp Val Val Val Gly Ser Cys Leu Ala Ile Gly Phe PheThr 305 310 315 320 gca gtc aaa tgg aga ggc aga tta ccc gta aat gat ctattt tgt agg 1307 Ala Val Lys Trp Arg Gly Arg Leu Pro Val Asn Asp Leu PheCys Arg 325 330 335 tgg gac cat gat acc ctg aaa ttc cat gac att tgg aacgag gac cca 1355 Trp Asp His Asp Thr Leu Lys Phe His Asp Ile Trp Asn GluAsp Pro 340 345 350 ctg agg gag aac tat ccc ttg gac ttg gag tct tca aaagac ttg gaa 1403 Leu Arg Glu Asn Tyr Pro Leu Asp Leu Glu Ser Ser Lys AspLeu Glu 355 360 365 aat cag ata cct ttg agt gat ttg aat tca tca tgt gatgaa caa cca 1451 Asn Gln Ile Pro Leu Ser Asp Leu Asn Ser Ser Cys Asp GluGln Pro 370 375 380 agt tca cca aag gga ttt aat agg aac cag aat aat aacttg ggt gat 1499 Ser Ser Pro Lys Gly Phe Asn Arg Asn Gln Asn Asn Asn LeuGly Asp 385 390 395 400 aag gga cag gga tta gaa caa gta gac gat caa ggagaa gaa tat gac 1547 Lys Gly Gln Gly Leu Glu Gln Val Asp Asp Gln Gly GluGlu Tyr Asp 405 410 415 aat tac gat ttt gat gat tct gat gac gac caa attgct gaa gct aat 1595 Asn Tyr Asp Phe Asp Asp Ser Asp Asp Asp Gln Ile AlaGlu Ala Asn 420 425 430 gat gtg gag aaa tcg gct aac aaa aca ata aag aagact cca aag tca 1643 Asp Val Glu Lys Ser Ala Asn Lys Thr Ile Lys Lys ThrPro Lys Ser 435 440 445 tac aag gaa tca ttc gaa gca aca tta tct aca aacttg aac tca caa 1691 Tyr Lys Glu Ser Phe Glu Ala Thr Leu Ser Thr Asn LeuAsn Ser Gln 450 455 460 att aca aat ttc cca tct tcc act cat ata cct cttaat tcc agt cgt 1739 Ile Thr Asn Phe Pro Ser Ser Thr His Ile Pro Leu AsnSer Ser Arg 465 470 475 480 tgacacaaac tcggttcttt aataactacc acttcgccctttacttagcg tctttgaaat 1799 tatagatctg gaattagaaa taaactacct ttttttttggccaattggaa aaatttaata 1859 caactcttat caagtcactc ctttattttc aatctttttctttatgtaca tgtaatttta 1919 cagagttcaa tagttaatta tttaattact tattttcactttatagttac actcttctcc 1979 ccacgtcctt ttttcttatc a 2000 5 480 PRTCandida kruisii 5 Met Leu Ser Leu Pro Ala Leu Ile Arg Arg Leu Tyr SerSer Phe Ile 1 5 10 15 Ser Ala Arg Glu Lys Gly Asn Ser Leu Pro Ala LeuLys Thr Ser Phe 20 25 30 Asn Pro Leu Pro Asn Phe Lys Thr Phe Pro Glu ArgLeu Val Ser Ala 35 40 45 Ser Ile Ser Asn Val Val His Phe Thr Ile Ile PheLeu Val Phe Val 50 55 60 Phe Ser Ile Ser Ile Met Pro Val Pro Leu Leu ValLys Ala Ile Leu 65 70 75 80 Ile Ile Val Leu Ile Tyr Ile Asn Phe Ala ProIle Phe Ala Gln Phe 85 90 95 Phe Asn Gly Gly Leu Pro Ile Leu Ser Trp ValIle Leu Phe Phe Asn 100 105 110 Ser Lys Asn Ile Pro Leu Ser Met Lys ArgPro Ile Ser Val Lys Val 115 120 125 Leu Pro Ala Leu Glu Thr Ile Phe TyrGly Asp Asn Leu Ser Glu Ile 130 135 140 Leu Ala Ser His Thr Asn Thr PheLeu Asp Leu Phe Ala Trp Phe Pro 145 150 155 160 Tyr Gly Ile Val His PheSer Leu Pro Phe Val Val Ala Ala Leu Ile 165 170 175 Phe Leu Phe Ala ProPro Lys Thr Leu Pro Gln Phe Cys Trp Ser Phe 180 185 190 Gly Tyr Met AsnLeu Val Gly Val Ile Ile Gln Asn Leu Val Phe Ser 195 200 205 Cys Ala ProPro Trp Tyr Lys Val Leu His Gly Leu Asp Lys Ala Asn 210 215 220 Tyr SerMet Lys Gly Ser Pro Gly Gly Leu Gly Arg Ile Asp Gly Leu 225 230 235 240Phe Gly Phe Asp Met Tyr Thr Ser Gly Phe Thr Asn Ser Pro Leu Ile 245 250255 Phe Gly Ala Met Pro Ser Leu His Ser Gly Cys Ser Thr Met Asp Ala 260265 270 Leu Trp Leu Ser Tyr Leu Phe Pro Lys Phe Thr Pro Leu Phe Val Phe275 280 285 Tyr Val Cys Trp Leu Trp Phe Ser Thr Met Tyr Leu Thr His HisTyr 290 295 300 Phe Ile Asp Val Val Val Gly Ser Cys Leu Ala Ile Gly PhePhe Thr 305 310 315 320 Ala Val Lys Trp Arg Gly Arg Leu Pro Val Asn AspLeu Phe Cys Arg 325 330 335 Trp Asp His Asp Thr Leu Lys Phe His Asp IleTrp Asn Glu Asp Pro 340 345 350 Leu Arg Glu Asn Tyr Pro Leu Asp Leu GluSer Ser Lys Asp Leu Glu 355 360 365 Asn Gln Ile Pro Leu Ser Asp Leu AsnSer Ser Cys Asp Glu Gln Pro 370 375 380 Ser Ser Pro Lys Gly Phe Asn ArgAsn Gln Asn Asn Asn Leu Gly Asp 385 390 395 400 Lys Gly Gln Gly Leu GluGln Val Asp Asp Gln Gly Glu Glu Tyr Asp 405 410 415 Asn Tyr Asp Phe AspAsp Ser Asp Asp Asp Gln Ile Ala Glu Ala Asn 420 425 430 Asp Val Glu LysSer Ala Asn Lys Thr Ile Lys Lys Thr Pro Lys Ser 435 440 445 Tyr Lys GluSer Phe Glu Ala Thr Leu Ser Thr Asn Leu Asn Ser Gln 450 455 460 Ile ThrAsn Phe Pro Ser Ser Thr His Ile Pro Leu Asn Ser Ser Arg 465 470 475 4806 2000 RNA Candida kruisii 6 acaguaaaaa gaaauugcgc ugcaugaaac uuuuuuuuuuuuuuuuuucg guguuucuuc 60 ucguuuuucu cguuuuucuc guuuuucucg guuucuuuugagaauguaau gaaauuugga 120 aacuguacaa acauucucag uguugauauu aacaauaauaaccacauuga cauugacaac 180 aucauuauua gguucuuucc auuuguauua uuuccaucucggacacguca gcuauagacc 240 gucgacacuc ccuauucccu gcaccaacua ggaagaacaggaagaacaag aagaacagaa 300 ugcucucgcu cccggcccuu auuaggcgcc uguauuccuccuuuauuucg gcaagagaaa 360 aggguaauuc auugccugca uuaaagacau ccuuuaacccccucccuaac uucaagaccu 420 ucccggagag acuggucucg gcauccauau ccaaugugguccauuucacg auuaucuuuc 480 uugucuuugu cuucuccauc ucaauaaugc cuguaccccuacuugugaaa gccauucuua 540 ucauugucuu gauuuacauc aacuuugcuc caauuuucgcccaguucuuu aacggcggcu 600 ugccgauccu cucauggguc auuuuguucu uuaacucaaagaacauaccu cugucaauga 660 agagacccau cuccgucaag gucuugcccg cauuggagacaaucuucuau ggcgauaauu 720 uaucugaaau cuuggcuucu cauacaaaca cuuucuuggacuuguuugcu ugguucccau 780 augguauagu ccauuucucc cugcccuuug uuguggcagcccuuauuuuu uuauuugccc 840 ccccuaaaac cuuaccccaa uuuuguuggu ccuuugguuauaugaauuug guuggcguca 900 uuauccaaaa cuugguuuuc ucaugugcuc caccaugguauaaagugcug cauggguugg 960 acaaggcaaa cuauucaaug aagggaucgc caggaggauuggguagaauc gauggacugu 1020 uugguuucga uauguacacc ucggguuuca ccaauucucccuugauuuuc ggugcaaugc 1080 ccuccuugca uucgggaugu ucaacuaugg acgcauuaugguugucuuau cuauucccua 1140 aauucacccc ucuauuuguu uucuacguuu gcugguuaugguucucuaca auguauuuga 1200 cucaucauua cuucaucgau guuguuguug guucuuguuuggcaauugga uucuucacug 1260 cagucaaaug gagaggcaga uuacccguaa augaucuauuuuguaggugg gaccaugaua 1320 cccugaaauu ccaugacauu uggaacgagg acccacugagggagaacuau cccuuggacu 1380 uggagucuuc aaaagacuug gaaaaucaga uaccuuugagugauuugaau ucaucaugug 1440 augaacaacc aaguucacca aagggauuua auaggaaccagaauaauaac uugggugaua 1500 agggacaggg auuagaacaa guagacgauc aaggagaagaauaugacaau uacgauuuug 1560 augauucuga ugacgaccaa auugcugaag cuaaugauguggagaaaucg gcuaacaaaa 1620 caauaaagaa gacuccaaag ucauacaagg aaucauucgaagcaacauua ucuacaaacu 1680 ugaacucaca aauuacaaau uucccaucuu ccacucauauaccucuuaau uccagucguu 1740 gacacaaacu cgguucuuua auaacuacca cuucgcccuuuacuuagcgu cuuugaaauu 1800 auagaucugg aauuagaaau aaacuaccuu uuuuuuuggccaauuggaaa aauuuaauac 1860 aacucuuauc aagucacucc uuuauuuuca aucuuuuucuuuauguacau guaauuuuac 1920 agaguucaau aguuaauuau uuaauuacuu auuuucacuuuauaguuaca cucuucuccc 1980 cacguccuuu uuucuuauca 2000 7 1949 DNA Candidaparapsilosis CDS (162)..(1679) 7 cacacacaca cacgggttag agcatctgaacttcacgaca actgttttct gtacaaccaa 60 aacaaaccac aacacaagac aacacaacacagtactttgc tgacattcaa taccaatacc 120 cgtctttttt ttttcaccac acaacgatattgctttgaag g atg tca cgc act tac 176 Met Ser Arg Thr Tyr 1 5 aat ggg tcttcc tcc tca tca ata ttt ctg aaa cca cgt cga ttt ttc 224 Asn Gly Ser SerSer Ser Ser Ile Phe Ser Lys Pro Arg Arg Phe Phe 10 15 20 aat aac tac tttcta ctg gaa aaa tca cca act tca aca tta cct gat 272 Asn Asn Tyr Phe LeuSer Glu Lys Ser Pro Thr Ser Thr Leu Pro Asp 25 30 35 tta gaa aca aac actagc atc aga act agt ctt cat aga cta aga cat 320 Leu Glu Thr Asn Thr SerIle Arg Thr Ser Leu His Arg Leu Arg His 40 45 50 cac aaa ttc ata aag ggagag att tta cac tat ggg ttt ctt tct ttt 368 His Lys Phe Ile Lys Gly GluIle Leu His Tyr Gly Phe Leu Ser Phe 55 60 65 gtt tgg tta ttt gtc ttt accata tac cca gca tcc atc ttg ttt aaa 416 Val Trp Leu Phe Val Phe Thr IleTyr Pro Ala Ser Ile Leu Phe Lys 70 75 80 85 att cca gtt tta ctt gca ttgcta acc att att act att cca gca act 464 Ile Pro Val Leu Leu Ala Leu LeuThr Ile Ile Thr Ile Pro Ala Thr 90 95 100 tca caa ttt ttt tta cca gcactt ccc att ttg act tgg ttg ggt ttt 512 Ser Gln Phe Phe Leu Pro Ala LeuPro Ile Leu Thr Trp Leu Gly Phe 105 110 115 tac ttc act agt tcc aaa atacct cat tct tgg aaa cca gct att tca 560 Tyr Phe Thr Ser Ser Lys Ile ProHis Ser Trp Lys Pro Ala Ile Ser 120 125 130 gtc aag gtt tta cca gcc atggag act atc ttg tat ggt gat gat tta 608 Val Lys Val Leu Pro Ala Met GluThr Ile Leu Tyr Gly Asp Asp Leu 135 140 145 tca aat gta ttg gca acc atcact aac cgt ggt ttg gat att ttg ggc 656 Ser Asn Val Leu Ala Thr Ile ThrAsn Arg Gly Leu Asp Ile Leu Gly 150 155 160 165 tgg tta cct tat ggt ttattt cat ttt ggt gct cca ttt gtt gta gct 704 Trp Leu Pro Tyr Gly Leu PheHis Phe Gly Ala Pro Phe Val Val Ala 170 175 180 gca att atg ttt ttc tttgct cca cca act agt tta aga tca ttt gcg 752 Ala Ile Met Phe Phe Phe AlaPro Pro Thr Ser Leu Arg Ser Phe Ala 185 190 195 ttt gct ttt ggt tat atgaac ttg ttt ggg gtt att att caa atg atg 800 Phe Ala Phe Gly Tyr Met AsnLeu Phe Gly Val Ile Ile Gln Met Met 200 205 210 ttt cca gct gct cca ccttgg tac aag aat ttg tat gga tta caa cca 848 Phe Pro Ala Ala Pro Pro TrpTyr Lys Asn Leu Tyr Gly Leu Gln Pro 215 220 225 gcc aac tat acc atg catggc tca cca ggt gga ttg ggc cgt ata gat 896 Ala Asn Tyr Thr Met His GlySer Pro Gly Gly Leu Gly Arg Ile Asp 230 235 240 245 gaa tta ttg gga gtcgat atg tac acc act ggg ttc tcc aac tca ccg 944 Glu Leu Leu Gly Val AspMet Tyr Thr Thr Gly Phe Ser Asn Ser Pro 250 255 260 gtt att ttt ggt gctttc cca tca tta cat tca ggt tgt tgt att atg 992 Val Ile Phe Gly Ala PhePro Ser Leu His Ser Gly Cys Cys Ile Met 265 270 275 gag gtt ttg ttt atgtgt tgg tta ttt ccc cgt ttg aaa gta ctt tgg 1040 Glu Val Leu Phe Met CysTrp Leu Phe Pro Arg Leu Lys Val Leu Trp 280 285 290 gtg ttt tat gca tcttgg tta tgg tgg agt aca atg tac ttg act cat 1088 Val Phe Tyr Ala Ser TrpLeu Trp Trp Ser Thr Met Tyr Leu Thr His 295 300 305 cac tat ttt gtt gattta act ggt ggt gct gta tta tca ttg gtt gtt 1136 His Tyr Phe Val Asp LeuThr Gly Gly Ala Val Leu Ser Leu Val Val 310 315 320 325 ttt gaa ttt gtcaag ttt aaa tac cta cct aag gct aac aag cgt tgt 1184 Phe Glu Phe Val LysPhe Lys Tyr Leu Pro Lys Ala Asn Lys Arg Cys 330 335 340 cgt tgg tct tatacg gaa ttg gaa tac tac agc aca acc aat gat gac 1232 Arg Trp Ser Tyr ThrGlu Leu Glu Tyr Tyr Ser Thr Thr Asn Asp Asp 345 350 355 cct tta gga ggtaac agt gct gcc act cga gtt gtt cgc ggc ggc gat 1280 Pro Leu Gly Gly AsnSer Ala Ala Thr Arg Val Val Arg Gly Gly Asp 360 365 370 ttg gaa aat tttgca tat tca aga gta tac cca tca ttg tca cag cag 1328 Leu Glu Asn Phe AlaTyr Ser Arg Val Tyr Pro Ser Leu Ser Gln Gln 375 380 385 caa caa ttg caagag cat ccg caa caa caa caa caa tca ctg tct aat 1376 Gln Gln Leu Gln GluHis Pro Gln Gln Gln Gln Gln Ser Ser Ser Asn 390 395 400 405 caa gaa aacata caa gag gag gtg ttt gaa atg ggc acc atc tca cgt 1424 Gln Glu Asn IleGln Glu Glu Val Phe Glu Met Gly Thr Ile Ser Arg 410 415 420 tca aga tcatca cca agg tcc ttt gtt tct ctt gga agt agc acc aat 1472 Ser Arg Ser SerPro Arg Ser Phe Val Ser Leu Gly Ser Ser Thr Asn 425 430 435 gtt gct gaagag aac ttt gac ctt gct gat gag gaa gaa gat atc ggt 1520 Val Ala Glu GluAsn Phe Asp Leu Ala Asp Glu Glu Glu Asp Ile Gly 440 445 450 tca gga tcatca aaa act ccc tca gta ttt gaa gaa gat aca caa aat 1568 Ser Gly Ser SerLys Thr Pro Ser Val Phe Glu Glu Asp Thr Gln Asn 455 460 465 tac gtc gtttct caa act acc tcc gtt gaa gac ttg act aat aat aca 1616 Tyr Val Val SerGln Thr Thr Ser Val Glu Asp Leu Thr Asn Asn Thr 470 475 480 485 caa gcatca gct gca ggc aac gga gct acc gat aag tca tca aga ctt 1664 Gln Ala SerAla Ala Gly Asn Gly Ala Thr Asp Lys Ser Ser Arg Leu 490 495 500 gga atcaat ggt tga acaaagggta gatgtattag aagagtaatc gtttcttttt 1719 Gly Ile AsnGly 505 taatggctgc atttgtctag aagagttcga ttataacgat gtttgatgatgtttatgttg 1779 atgattcacg atttttacga attgtacaag aaccctattc acaacctgagatgggattag 1839 aatgtattga acttggaaaa gatatgggac gtgtgcttgg aatgtcctccttttctcata 1899 tccactgccc cattgctatc cctttacaag attattacca tgttatgtct1949 8 505 PRT Candida parapsilosis 8 Met Ser Arg Thr Tyr Asn Gly SerSer Ser Ser Ser Ile Phe Ser Lys 1 5 10 15 Pro Arg Arg Phe Phe Asn AsnTyr Phe Leu Ser Glu Lys Ser Pro Thr 20 25 30 Ser Thr Leu Pro Asp Leu GluThr Asn Thr Ser Ile Arg Thr Ser Leu 35 40 45 His Arg Leu Arg His His LysPhe Ile Lys Gly Glu Ile Leu His Tyr 50 55 60 Gly Phe Leu Ser Phe Val TrpLeu Phe Val Phe Thr Ile Tyr Pro Ala 65 70 75 80 Ser Ile Leu Phe Lys IlePro Val Leu Leu Ala Leu Leu Thr Ile Ile 85 90 95 Thr Ile Pro Ala Thr SerGln Phe Phe Leu Pro Ala Leu Pro Ile Leu 100 105 110 Thr Trp Leu Gly PheTyr Phe Thr Ser Ser Lys Ile Pro His Ser Trp 115 120 125 Lys Pro Ala IleSer Val Lys Val Leu Pro Ala Met Glu Thr Ile Leu 130 135 140 Tyr Gly AspAsp Leu Ser Asn Val Leu Ala Thr Ile Thr Asn Arg Gly 145 150 155 160 LeuAsp Ile Leu Gly Trp Leu Pro Tyr Gly Leu Phe His Phe Gly Ala 165 170 175Pro Phe Val Val Ala Ala Ile Met Phe Phe Phe Ala Pro Pro Thr Ser 180 185190 Leu Arg Ser Phe Ala Phe Ala Phe Gly Tyr Met Asn Leu Phe Gly Val 195200 205 Ile Ile Gln Met Met Phe Pro Ala Ala Pro Pro Trp Tyr Lys Asn Leu210 215 220 Tyr Gly Leu Gln Pro Ala Asn Tyr Thr Met His Gly Ser Pro GlyGly 225 230 235 240 Leu Gly Arg Ile Asp Glu Leu Leu Gly Val Asp Met TyrThr Thr Gly 245 250 255 Phe Ser Asn Ser Pro Val Ile Phe Gly Ala Phe ProSer Leu His Ser 260 265 270 Gly Cys Cys Ile Met Glu Val Leu Phe Met CysTrp Leu Phe Pro Arg 275 280 285 Leu Lys Val Leu Trp Val Phe Tyr Ala SerTrp Leu Trp Trp Ser Thr 290 295 300 Met Tyr Leu Thr His His Tyr Phe ValAsp Leu Thr Gly Gly Ala Val 305 310 315 320 Leu Ser Leu Val Val Phe GluPhe Val Lys Phe Lys Tyr Leu Pro Lys 325 330 335 Ala Asn Lys Arg Cys ArgTrp Ser Tyr Thr Glu Leu Glu Tyr Tyr Ser 340 345 350 Thr Thr Asn Asp AspPro Leu Gly Gly Asn Ser Ala Ala Thr Arg Val 355 360 365 Val Arg Gly GlyAsp Leu Glu Asn Phe Ala Tyr Ser Arg Val Tyr Pro 370 375 380 Ser Leu SerGln Gln Gln Gln Leu Gln Glu His Pro Gln Gln Gln Gln 385 390 395 400 GlnSer Ser Ser Asn Gln Glu Asn Ile Gln Glu Glu Val Phe Glu Met 405 410 415Gly Thr Ile Ser Arg Ser Arg Ser Ser Pro Arg Ser Phe Val Ser Leu 420 425430 Gly Ser Ser Thr Asn Val Ala Glu Glu Asn Phe Asp Leu Ala Asp Glu 435440 445 Glu Glu Asp Ile Gly Ser Gly Ser Ser Lys Thr Pro Ser Val Phe Glu450 455 460 Glu Asp Thr Gln Asn Tyr Val Val Ser Gln Thr Thr Ser Val GluAsp 465 470 475 480 Leu Thr Asn Asn Thr Gln Ala Ser Ala Ala Gly Asn GlyAla Thr Asp 485 490 495 Lys Ser Ser Arg Leu Gly Ile Asn Gly 500 505 91788 RNA Candida parapsilosis 9 augucacgca cuuacaaugg gucuuccuccucaucaauau uucugaaacc acgucgauuu 60 uucaauaacu acuuucuacu ggaaaaaucaccaacuucaa cauuaccuga uuuagaaaca 120 aacacuagca ucagaacuag ucuucauagacuaagacauc acaaauucau aaagggagag 180 auuuuacacu auggguuucu uucuuuuguuugguuauuug ucuuuaccau auacccagca 240 uccaucuugu uuaaaauucc aguuuuacuugcauugcuaa ccauuauuac uauuccagca 300 acuucacaau uuuuuuuacc agcacuucccauuuugacuu gguuggguuu uuacuucacu 360 aguuccaaaa uaccucauuc uuggaaaccagcuauuucag ucaagguuuu accagccaug 420 gagacuaucu uguaugguga ugauuuaucaaauguauugg caaccaucac uaaccguggu 480 uuggauauuu ugggcugguu accuuaugguuuauuucauu uuggugcucc auuuguugua 540 gcugcaauua uguuuuucuu ugcuccaccaacuaguuuaa gaucauuugc guuugcuuuu 600 gguuauauga acuuguuugg gguuauuauucaaaugaugu uuccagcugc uccaccuugg 660 uacaagaauu uguauggauu acaaccagccaacuauacca ugcauggcuc accaggugga 720 uugggccgua uagaugaauu auugggagucgauauguaca ccacuggguu cuccaacuca 780 ccgguuauuu uuggugcuuu cccaucauuacauucagguu guuguauuau ggagguuuug 840 uuuauguguu gguuauuucc ccguuugaaaguacuuuggg uguuuuaugc aucuugguua 900 ugguggagua caauguacuu gacucaucacuauuuuguug auuuaacugg uggugcugua 960 uuaucauugg uuguuuuuga auuugucaaguuuaaauacc uaccuaaggc uaacaagcgu 1020 ugucguuggu cuuauacgga auuggaauacuacagcacaa ccaaugauga cccuuuagga 1080 gguaacagug cugccacucg aguuguucgcggcggcgauu uggaaaauuu ugcauauuca 1140 agaguauacc caucauuguc acagcagcaacaauugcaag agcauccgca acaacaacaa 1200 caaucacugu cuaaucaaga aaacauacaagaggaggugu uugaaauggg caccaucuca 1260 cguucaagau caucaccaag guccuuuguuucucuuggaa guagcaccaa uguugcugaa 1320 gagaacuuug accuugcuga ugaggaagaagauaucgguu caggaucauc aaaaacuccc 1380 ucaguauuug aagaagauac acaaaauuacgucguuucuc aaacuaccuc cguugaagac 1440 uugacuaaua auacacaagc aucagcugcaggcaacggag cuaccgauaa gucaucaaga 1500 cuuggaauca augguugaac aaaggguagauguauuagaa gaguaaucgu uucuuuuuua 1560 auggcugcau uugucuagaa gaguucgauuauaacgaugu uugaugaugu uuauguugau 1620 gauucacgau uuuuacgaau uguacaagaacccuauucac aaccugagau gggauuagaa 1680 uguauugaac uuggaaaaga uaugggacgugugcuuggaa uguccuccuu uucucauauc 1740 cacugcccca uugcuauccc uuuacaagauuauuaccaug uuaugucu 1788 10 2214 DNA Candida tropicalis CDS(601)..(1974) 10 cgatggtgat ttctttttct ctcctattcc tgtaccccgc aaatgtgtgatttcaatccc 60 aaggtgccaa agcacactga tgttggcttg aaagactcag atatggagctttgtaatata 120 agaaaaatgc caaagtgctt ttcttgccaa tcattgtcga tttggacttttctgcctctg 180 tctcactttg catctttagt gacgcatacg ggaaaaaatg taaaatgcgctgaccataaa 240 tgacattcgg aacataatgc atagaatatt aataccgcat accgataacagaagtcacaa 300 acgtcacaaa acgtttcaaa aaaacagaaa caggcaagcc tgaaagttgaacacagccac 360 ttcttcccca aactatgaaa acttggtcgt tgcaagatca ttcacttgcgcccctatttt 420 atagcatgga aaggatgcgt ttctgttccg gcccatttgc gcgcatttgaggtcacgtgc 480 gctgcatcgt cacggcccat tcctttttct atccttttat tttatgttgcttcagcccca 540 acgctgaaag gccgtctttt ccagcagaat ccattactct gctatatttccctcattcca 600 atg tca acg gtc cta agg ttc aag cct ttc cgc gtt gct tacaag caa 648 Met Ser Thr Val Leu Arg Phe Lys Pro Phe Arg Val Ala Tyr LysGln 1 5 10 15 ttc cac aat tat ttt ttg ctg gag aaa gaa gca gga tgc actttg gcg 696 Phe His Asn Tyr Phe Leu Leu Glu Lys Glu Ala Gly Cys Thr LeuAla 20 25 30 gat ctt gat ttc gat act gat atc cgc aag tcc atc cag aaa ctacgt 744 Asp Leu Asp Phe Asp Thr Asp Ile Arg Lys Ser Ile Gln Lys Leu Arg35 40 45 tcc cat gag ttt acg ccg ggc gaa ata gtg cac tac act ttc ctc acg792 Ser His Glu Phe Thr Pro Gly Glu Ile Val His Tyr Thr Phe Leu Thr 5055 60 ctg ttg atc ttg ttt gtt tat gtc att ttc ccc gcg tcg ttt ttg ctc840 Ser Leu Ile Leu Phe Val Tyr Val Ile Phe Pro Ala Ser Phe Leu Leu 6570 75 80 aaa acg ccc atc ttg gcg gct ttc ggc ttt tgt ttc tta ttg ccc ttg888 Lys Thr Pro Ile Leu Ala Ala Phe Gly Phe Cys Phe Leu Leu Pro Leu 8590 95 acg tcc cag ttt ttc gtg tca gcg ttg ccc att ttc aca tgg ttg gcg936 Thr Ser Gln Phe Phe Val Ser Ala Leu Pro Ile Phe Thr Trp Leu Ala 100105 110 ctt ttc ttc tct gcc agc aag atc cct gtg gac tgg aag cct gtc att984 Leu Phe Phe Ser Ala Ser Lys Ile Pro Val Asp Trp Lys Pro Val Ile 115120 125 agc gtc aag ttt ctt cct gcc atg gaa act atc ttg tac gga gac aac1032 Ser Val Lys Phe Leu Pro Ala Met Glu Thr Ile Leu Tyr Gly Asp Asn 130135 140 ttg tcg aat gtt ttg gca gcc acc aac aac tgg gtg cta gat atc ttg1080 Leu Ser Asn Val Leu Ala Ala Thr Asn Asn Trp Val Leu Asp Ile Leu 145150 155 160 gcg tgg ttg ccc tac ggt atc atc cac ttt ctg tct cca ttt gtagtt 1128 Ala Trp Leu Pro Tyr Gly Ile Ile His Phe Ser Ser Pro Phe Val Val165 170 175 gcg ctc ttc att ttc ctt ttt gct ccg ccc act tct ttg cgc tcgttt 1176 Ala Leu Phe Ile Phe Leu Phe Ala Pro Pro Thr Ser Leu Arg Ser Phe180 185 190 ggc ttc gcc ttt ggc tac atg aat gtg gtt ggt gtg ttg atc cagctt 1224 Gly Phe Ala Phe Gly Tyr Met Asn Val Val Gly Val Leu Ile Gln Leu195 200 205 tgt ttc ccc tca gct gct ccg tgg tac aag aac ttg cac ggc ctagaa 1272 Cys Phe Pro Ser Ala Ala Pro Trp Tyr Lys Asn Leu His Gly Leu Glu210 215 220 cct gca aat tat tcc atg aac ggg tct cct ggt ggc ttg ggc cgcatt 1320 Pro Ala Asn Tyr Ser Met Asn Gly Ser Pro Gly Gly Leu Gly Arg Ile225 230 235 240 gac gag ctt ttc ggt gtc gac atg tac acc act act ttc atgaac tca 1368 Asp Glu Leu Phe Gly Val Asp Met Tyr Thr Thr Thr Phe Met AsnSer 245 250 255 cca ttg gtt ttc ggc gcg ttc ccc tcg ctc cat tcg ggc tgcgca gtc 1416 Pro Leu Val Phe Gly Ala Phe Pro Ser Leu His Ser Gly Cys AlaVal 260 265 270 atg gat gtg ctc ttt ttg tgc tgg ctc ttc cct cgt ttc acctat gtt 1464 Met Asp Val Leu Phe Leu Cys Trp Leu Phe Pro Arg Phe Thr TyrVal 275 280 285 tgg tgg agc tac gcc tgt ctt ttg tgg tgg agc acc atg tatttg acg 1512 Trp Trp Ser Tyr Ala Cys Leu Leu Trp Trp Ser Thr Met Tyr LeuThr 290 295 300 cac cac tat ttt atc gac ttg atc ttc ggt gcc gct ttg tctgtg cta 1560 His His Tyr Phe Ile Asp Leu Ile Phe Gly Ala Ala Leu Ser ValLeu 305 310 315 320 gtt ttc aac tat gtc aag tac acg cag ttg cca gtg gtggac ccc acc 1608 Val Phe Asn Tyr Val Lys Tyr Thr Gln Leu Pro Val Val AspPro Thr 325 330 335 aag ttc tgt cgc tgg ctg tac acc acc gtg gag ctt cttgac att aac 1656 Lys Phe Cys Arg Trp Ser Tyr Thr Thr Val Glu Leu Leu AspIle Asn 340 345 350 gcc aac gac cca ttg cag agt ttc gtg cca tta cac ggagac atg gag 1704 Ala Asn Asp Pro Leu Gln Ser Phe Val Pro Leu His Gly AspMet Glu 355 360 365 gaa ggt atg ctg cat ccc ttt tac cac gcg cca gcg cttgag aat ttc 1752 Glu Gly Met Ser His Pro Phe Tyr His Ala Pro Ala Leu GluAsn Phe 370 375 380 gaa atg tct act ata aac cgt tca cgg gaa agc gtg gctgcg ccg ttt 1800 Glu Met Ser Thr Ile Asn Arg Ser Arg Glu Ser Val Ala AlaPro Phe 385 390 395 400 ccg acc ata gcc gag gat gtg gct gag gaa acc gacagc agc gcg tcc 1848 Pro Thr Ile Ala Glu Asp Val Ala Glu Glu Thr Asp SerSer Ala Ser 405 410 415 aat tcg gtt ttt gat gct gag cgt ttt gac gaa gaaacg cat cct agc 1896 Asn Ser Val Phe Asp Ala Glu Arg Phe Asp Glu Glu ThrHis Pro Ser 420 425 430 ctg cgt acg tcc agc acc tcg ttg aac gac ttg cttctt ccc ctg agc 1944 Ser Arg Thr Ser Ser Thr Ser Leu Asn Asp Leu Leu LeuPro Ser Ser 435 440 445 tcg agc agc tca aag ccc aaa aga agc acgtagcctttta cattgtcgca 1994 Ser Ser Ser Ser Lys Pro Lys Arg Ser Thr 450455 ccgctctttg gccaggagca ctttaatatt ggccgtgtat actacataaa tgcgacgtct2054 atcgtgatgc tatgtacatt gtgagtccta gctcggtata tgatatatcc tctttcgttt2114 tcgttcgtag cttccttcac ttcctaatcg ttccttctcc ttagtttgac accttctcga2174 acactctttg gtaagtagtg cccacattca agttgccctt 2214 11 458 PRT Candidatropicalis 11 Met Ser Thr Val Leu Arg Phe Lys Pro Phe Arg Val Ala TyrLys Gln 1 5 10 15 Phe His Asn Tyr Phe Leu Ser Glu Lys Glu Ala Gly CysThr Leu Ala 20 25 30 Asp Leu Asp Phe Asp Thr Asp Ile Arg Lys Ser Ile GlnLys Leu Arg 35 40 45 Ser His Glu Phe Thr Pro Gly Glu Ile Val His Tyr ThrPhe Leu Thr 50 55 60 Leu Ser Ile Leu Phe Val Tyr Val Ile Phe Pro Ala SerPhe Leu Leu 65 70 75 80 Lys Thr Pro Ile Leu Ala Ala Phe Gly Phe Cys PheLeu Leu Pro Leu 85 90 95 Thr Ser Gln Phe Phe Val Ser Ala Leu Pro Ile PheThr Trp Leu Ala 100 105 110 Leu Phe Phe Ser Ala Ser Lys Ile Pro Val AspTrp Lys Pro Val Ile 115 120 125 Ser Val Lys Phe Leu Pro Ala Met Glu ThrIle Leu Tyr Gly Asp Asn 130 135 140 Leu Ser Asn Val Leu Ala Ala Thr AsnAsn Trp Val Leu Asp Ile Leu 145 150 155 160 Ala Trp Leu Pro Tyr Gly IleIle His Phe Ser Ser Pro Phe Val Val 165 170 175 Ala Leu Phe Ile Phe LeuPhe Ala Pro Pro Thr Ser Leu Arg Ser Phe 180 185 190 Gly Phe Ala Phe GlyTyr Met Asn Val Val Gly Val Leu Ile Gln Leu 195 200 205 Cys Phe Pro SerAla Ala Pro Trp Tyr Lys Asn Leu His Gly Leu Glu 210 215 220 Pro Ala AsnTyr Ser Met Asn Gly Ser Pro Gly Gly Leu Gly Arg Ile 225 230 235 240 AspGlu Leu Phe Gly Val Asp Met Tyr Thr Thr Thr Phe Met Asn Ser 245 250 255Pro Leu Val Phe Gly Ala Phe Pro Ser Leu His Ser Gly Cys Ala Val 260 265270 Met Asp Val Leu Phe Leu Cys Trp Leu Phe Pro Arg Phe Thr Tyr Val 275280 285 Trp Trp Ser Tyr Ala Cys Leu Leu Trp Trp Ser Thr Met Tyr Leu Thr290 295 300 His His Tyr Phe Ile Asp Leu Ile Phe Gly Ala Ala Leu Ser ValLeu 305 310 315 320 Val Phe Asn Tyr Val Lys Tyr Thr Gln Leu Pro Val ValAsp Pro Thr 325 330 335 Lys Phe Cys Arg Trp Ser Tyr Thr Thr Val Glu LeuLeu Asp Ile Asn 340 345 350 Ala Asn Asp Pro Leu Gln Ser Phe Val Pro LeuHis Gly Asp Met Glu 355 360 365 Glu Gly Met Ser His Pro Phe Tyr His AlaPro Ala Leu Glu Asn Phe 370 375 380 Glu Met Ser Thr Ile Asn Arg Ser ArgGlu Ser Val Ala Ala Pro Phe 385 390 395 400 Pro Thr Ile Ala Glu Asp ValAla Glu Glu Thr Asp Ser Ser Ala Ser 405 410 415 Asn Ser Val Phe Asp AlaGlu Arg Phe Asp Glu Glu Thr His Pro Ser 420 425 430 Ser Arg Thr Ser SerThr Ser Leu Asn Asp Leu Leu Leu Pro Ser Ser 435 440 445 Ser Ser Ser SerLys Pro Lys Arg Ser Thr 450 455 12 2214 RNA Candida tropicalis 12cgauggugau uucuuuuucu cuccuauucc uguaccccgc aaauguguga uuucaauccc 60aaggugccaa agcacacuga uguuggcuug aaagacucag auauggagcu uuguaauaua 120agaaaaaugc caaagugcuu uucuugccaa ucauugucga uuuggacuuu ucugccucug 180ucucacuuug caucuuuagu gacgcauacg ggaaaaaaug uaaaaugcgc ugaccauaaa 240ugacauucgg aacauaaugc auagaauauu aauaccgcau accgauaaca gaagucacaa 300acgucacaaa acguuucaaa aaaacagaaa caggcaagcc ugaaaguuga acacagccac 360uucuucccca aacuaugaaa acuuggucgu ugcaagauca uucacuugcg ccccuauuuu 420auagcaugga aaggaugcgu uucuguuccg gcccauuugc gcgcauuuga ggucacgugc 480gcugcaucgu cacggcccau uccuuuuucu auccuuuuau uuuauguugc uucagcccca 540acgcugaaag gccgucuuuu ccagcagaau ccauuacucu gcuauauuuc ccucauucca 600augucaacgg uccuaagguu caagccuuuc cgcguugcuu acaagcaauu ccacaauuau 660uuuuugcugg agaaagaagc aggaugcacu uuggcggauc uugauuucga uacugauauc 720cgcaagucca uccagaaacu acguucccau gaguuuacgc cgggcgaaau agugcacuac 780acuuuccuca cgcuguugau cuuguuuguu uaugucauuu uccccgcguc guuuuugcuc 840aaaacgccca ucuuggcggc uuucggcuuu uguuucuuau ugcccuugac gucccaguuu 900uucgugucag cguugcccau uuucacaugg uuggcgcuuu ucuucucugc cagcaagauc 960ccuguggacu ggaagccugu cauuagcguc aaguuucuuc cugccaugga aacuaucuug 1020uacggagaca acuugucgaa uguuuuggca gccaccaaca acugggugcu agauaucuug 1080gcgugguugc ccuacgguau cauccacuuu cugucuccau uuguaguugc gcucuucauu 1140uuccuuuuug cuccgcccac uucuuugcgc ucguuuggcu ucgccuuugg cuacaugaau 1200gugguuggug uguugaucca gcuuuguuuc cccucagcug cuccguggua caagaacuug 1260cacggccuag aaccugcaaa uuauuccaug aacgggucuc cugguggcuu gggccgcauu 1320gacgagcuuu ucggugucga cauguacacc acuacuuuca ugaacucacc auugguuuuc 1380ggcgcguucc ccucgcucca uucgggcugc gcagucaugg augugcucuu uuugugcugg 1440cucuucccuc guuucaccua uguuuggugg agcuacgccu gucuuuugug guggagcacc 1500auguauuuga cgcaccacua uuuuaucgac uugaucuucg gugccgcuuu gucugugcua 1560guuuucaacu augucaagua cacgcaguug ccaguggugg accccaccaa guucugucgc 1620uggcuguaca ccaccgugga gcuucuugac auuaacgcca acgacccauu gcagaguuuc 1680gugccauuac acggagacau ggaggaaggu augcugcauc ccuuuuacca cgcgccagcg 1740cuugagaauu ucgaaauguc uacuauaaac cguucacggg aaagcguggc ugcgccguuu 1800ccgaccauag ccgaggaugu ggcugaggaa accgacagca gcgcguccaa uucgguuuuu 1860gaugcugagc guuuugacga agaaacgcau ccuagccugc guacguccag caccucguug 1920aacgacuugc uucuuccccu gagcucgagc agcucaaagc ccaaaagaag cacguagccu 1980uuuacauugu cgcaccgcuc uuuggccagg agcacuuuaa uauuggccgu guauacuaca 2040uaaaugcgac gucuaucgug augcuaugua cauugugagu ccuagcucgg uauaugauau 2100auccucuuuc guuuucguuc guagcuuccu ucacuuccua aucguuccuu cuccuuaguu 2160ugacaccuuc ucgaacacuc uuugguaagu agugcccaca uucaaguugc ccuu 2214 13 2935DNA Aspergillus fumigatus CDS (450)..(1757) 13 attttcttcc ccataacaactcttctcgcc cttcctccgg ctccgtggcc aaattgtttt 60 atgcagcgcc tcctagcgatttaacctcgt tctcgttgcc cttgcctgtc cgccttgcgt 120 cagtacgacc cttgcaacgtgaccttcccc agagtatcct cgtttggccg ctggagaccg 180 gagcttgcac cctcataaactagctcttcg aaatcaattc tccgttctcc agagattatc 240 ggatcgaatc tctccgctgtcgacaccttt cgtctctcgg tgatcctcgc ccttggagtc 300 tcgtcacgtt gacgccttgaacccctggcc gccaactcca cataggagac cacacttcat 360 tcttcccccg ccataattgcagcaccctcc gtctcccttc gagctcctcc tggatcatca 420 agtccgaaag gattagactcgtcgcagcg atg aat acc acc ctt cca tcc tgg 473 Met Asn Thr Thr Leu ProSer Trp 1 5 aag gat cgg acg caa aac cag ttc ggc aag ctc cag atc caa gtccca 521 Lys Asp Arg Thr Gln Asn Gln Phe Gly Lys Leu Gln Ile Gln Val Pro10 15 20 tgg cgc acc ata caa ctt ctc gtg ccg cac cgt atg cga cgg aag att569 Trp Arg Thr Ile Gln Leu Leu Val Pro His Arg Met Arg Arg Lys Ile 2530 35 40 cgg tcc aag ctg cgc agt cgg atc tcg cct acc tca tcg ata tcc tcg617 Arg Ser Lys Leu Arg Ser Arg Ile Ser Pro Thr Ser Ser Ile Ser Ser 4550 55 ttg cag acg tca ttc tca cct gtc gat aca ctc agg tcg ctg caa agt665 Leu Gln Thr Ser Phe Ser Pro Val Asp Thr Leu Arg Ser Leu Gln Ser 6065 70 cat aga tgg acg ctc tat gac ttt cag tat ctt ttg ctg ctg att gtc713 His Arg Trp Thr Leu Tyr Asp Phe Gln Tyr Leu Leu Leu Leu Ile Val 7580 85 ggc ata ttc tcg ctg agc gtt atg gaa tca cct gga cca ttg gca aag761 Gly Ile Phe Ser Leu Ser Val Met Glu Ser Pro Gly Pro Leu Ala Lys 9095 100 acc gcc gcg ttt acg cta ctt ctc gtc tct ctc ctt ctc ccg att acg809 Thr Ala Ala Phe Thr Leu Leu Leu Val Ser Leu Leu Leu Pro Ile Thr 105110 115 120 cgc cag ttc ttc ttg cca ttc ctc ccg att gca gga tgg ctt atattt 857 Arg Gln Phe Phe Leu Pro Phe Leu Pro Ile Ala Gly Trp Leu Ile Phe125 130 135 ttc tac gct tgc cag ttc atc ccg agc gac tgg cgc cct gca atctgg 905 Phe Tyr Ala Cys Gln Phe Ile Pro Ser Asp Trp Arg Pro Ala Ile Trp140 145 150 gtt cgc gtg ctg ccg gct ctg gaa aac att ctc tac ggt gct aatatc 953 Val Arg Val Leu Pro Ala Leu Glu Asn Ile Leu Tyr Gly Ala Asn Ile155 160 165 agt aac atc ctt tcc gct cac caa aat gtg gtg ctt gac gtt ttggcg 1001 Ser Asn Ile Leu Ser Ala His Gln Asn Val Val Leu Asp Val Leu Ala170 175 180 tgg ctt ccc tac gga atc tgc cat tat ggc gcg cca ttt gtg tgctca 1049 Trp Leu Pro Tyr Gly Ile Cys His Tyr Gly Ala Pro Phe Val Cys Ser185 190 195 200 gcg atc atg ttc atc ttt ggt cct ccc ggc acc gtc ccc cttttc gct 1097 Ala Ile Met Phe Ile Phe Gly Pro Pro Gly Thr Val Pro Leu PheAla 205 210 215 cga act ttt gga tac atc agc atg gct gca gtc acc att cagctg ttt 1145 Arg Thr Phe Gly Tyr Ile Ser Met Ala Ala Val Thr Ile Gln LeuPhe 220 225 230 ttc ccc tgc tct cct ccg tgg tac gaa aat ctg tat ggt ttggct ccg 1193 Phe Pro Cys Ser Pro Pro Trp Tyr Glu Asn Leu Tyr Gly Leu AlaPro 235 240 245 gct gat tac tcc atg ccg ggt aat cct gcg ggc ctt gct cgcatc gat 1241 Ala Asp Tyr Ser Met Pro Gly Asn Pro Ala Gly Leu Ala Arg IleAsp 250 255 260 gag ctt ttt ggg ata gac ttg tac aca tcg ggc ttc aga caatct ccc 1289 Glu Leu Phe Gly Ile Asp Leu Tyr Thr Ser Gly Phe Arg Gln SerPro 265 270 275 280 gtc gtg ttt ggc gca ttt cct tcc cta cat gcc gct gattcg aca ctt 1337 Val Val Phe Gly Ala Phe Pro Ser Leu His Ala Ala Asp SerThr Leu 285 290 295 gca gct cta ttt atg agc caa gtg ttc cca cgg ttg aagccc ttg ttt 1385 Ala Ala Leu Phe Met Ser Gln Val Phe Pro Arg Leu Lys ProLeu Phe 300 305 310 gtc atc tat act ctc tgg atg tgg tgg gct aca atg tatctt tcg cac 1433 Val Ile Tyr Thr Leu Trp Met Trp Trp Ala Thr Met Tyr LeuSer His 315 320 325 cac tac gct gtt gat ctg gtc ggt ggt ggc ctc ttg gcaact gtc gcg 1481 His Tyr Ala Val Asp Leu Val Gly Gly Gly Leu Leu Ala ThrVal Ala 330 335 340 ttc tac ttt gct aaa acg cgg ttc atg cct cgc gtc cagaat gat aag 1529 Phe Tyr Phe Ala Lys Thr Arg Phe Met Pro Arg Val Gln AsnAsp Lys 345 350 355 360 atg ttc cgc tgg gac tac gat tat gtt gag tca ggcgat tcc gca ctc 1577 Met Phe Arg Trp Asp Tyr Asp Tyr Val Glu Ser Gly AspSer Ala Leu 365 370 375 gac tat ggg tac ggt cca gcc agc ttc gaa ggc gaattc aac ctt gat 1625 Asp Tyr Gly Tyr Gly Pro Ala Ser Phe Glu Gly Glu PheAsn Leu Asp 380 385 390 agc gat gag tgg acc gtt ggt tct tcg tca tcc atttcg tcc ggc tcc 1673 Ser Asp Glu Trp Thr Val Gly Ser Ser Ser Ser Ile SerSer Gly Ser 395 400 405 ctc agt cca gtt gac gac cac tac tct tgg gag ggcgag act ctt gcc 1721 Leu Ser Pro Val Asp Asp His Tyr Ser Trp Glu Gly GluThr Leu Ala 410 415 420 tct cct gcc acc gag atc gag tct gga agg cat ttctgatcctgct 1767 Ser Pro Ala Thr Glu Ile Glu Ser Gly Arg His Phe 425 430435 caatgagcct tgatacgtac tacactgtgt acgtgctact gcattgacta atgagacggc1827 gttttcaaac aaattttaac gacatgcttg gttatcgcat tgagctgatt tcgacacata1887 tatatgttta atacgttttg gggacactcc agggattcat gacggttgct tcatatcccg1947 acctggggat ggattgacct ggttgtgccc aattttcttc tgcctaacgt tttgattata2007 catgcatttt tcacgaaacc agccggcccg ccatgatcgt gacctcaatt tgagctcgaa2067 tcttcctggg gcctccagcg ataattctta atgctcgttc cgagggtgcc acatcggaca2127 ttcgcttgta caacttttgc agaacgaaca ttttcaccga ttccaacttg agtcattcgc2187 ttactacttt caactggtcg agaaacttcg cttcttttca gctcggctag gtgcataaat2247 attttacatt cgtgtcgatc gctcacattt cacggcgcct ggaaacttgg gggtttcgat2307 ttcattggaa aggataaaca acatgggctg ggcgcctttt acacgcacta cattcgctta2367 gaaaagttct gatgctttta atgattcttg cattggcata tagaaagggg tcctccagac2427 tcgctactgt ggtcctctct caaccccaca ctcgcttgct tttaacagtg gacaccccgt2487 ggagctacgt ctccatcaaa tatttggcat caaccggaat cgatgccagg aggactgagc2547 ttactcacgg tgaccgtcgg gtaaaaggcg ttctaacaga acatctcctc tatcctcctg2607 tcccatctcg attctctggc tgctggacgc aacacacctc gctgcacgtt ttcgacttcc2667 taatacgacc taatatcatt ctcggttttc tttgctctgg ctcgcggccg ccatttatat2727 ggcgtgtcgg ctcgagtctg aagtgaactt ttttctcctt tctggcctcc acaactttcc2787 gatccctagc agcttcccgt gcacagcgag gtgttgttgg atgattgttc catagcatta2847 tcattattcc taatccggta gcgttatgat ttatgaagaa cagtgatgta cattattatg2907 cggtgactaa aaaaaaaaaa aaaaaaaa 2935 14 436 PRT Aspergillusfumigatus 14 Met Asn Thr Thr Leu Pro Ser Trp Lys Asp Arg Thr Gln Asn GlnPhe 1 5 10 15 Gly Lys Leu Gln Ile Gln Val Pro Trp Arg Thr Ile Gln LeuLeu Val 20 25 30 Pro His Arg Met Arg Arg Lys Ile Arg Ser Lys Leu Arg SerArg Ile 35 40 45 Ser Pro Thr Ser Ser Ile Ser Ser Leu Gln Thr Ser Phe SerPro Val 50 55 60 Asp Thr Leu Arg Ser Leu Gln Ser His Arg Trp Thr Leu TyrAsp Phe 65 70 75 80 Gln Tyr Leu Leu Leu Leu Ile Val Gly Ile Phe Ser LeuSer Val Met 85 90 95 Glu Ser Pro Gly Pro Leu Ala Lys Thr Ala Ala Phe ThrLeu Leu Leu 100 105 110 Val Ser Leu Leu Leu Pro Ile Thr Arg Gln Phe PheLeu Pro Phe Leu 115 120 125 Pro Ile Ala Gly Trp Leu Ile Phe Phe Tyr AlaCys Gln Phe Ile Pro 130 135 140 Ser Asp Trp Arg Pro Ala Ile Trp Val ArgVal Leu Pro Ala Leu Glu 145 150 155 160 Asn Ile Leu Tyr Gly Ala Asn IleSer Asn Ile Leu Ser Ala His Gln 165 170 175 Asn Val Val Leu Asp Val LeuAla Trp Leu Pro Tyr Gly Ile Cys His 180 185 190 Tyr Gly Ala Pro Phe ValCys Ser Ala Ile Met Phe Ile Phe Gly Pro 195 200 205 Pro Gly Thr Val ProLeu Phe Ala Arg Thr Phe Gly Tyr Ile Ser Met 210 215 220 Ala Ala Val ThrIle Gln Leu Phe Phe Pro Cys Ser Pro Pro Trp Tyr 225 230 235 240 Glu AsnLeu Tyr Gly Leu Ala Pro Ala Asp Tyr Ser Met Pro Gly Asn 245 250 255 ProAla Gly Leu Ala Arg Ile Asp Glu Leu Phe Gly Ile Asp Leu Tyr 260 265 270Thr Ser Gly Phe Arg Gln Ser Pro Val Val Phe Gly Ala Phe Pro Ser 275 280285 Leu His Ala Ala Asp Ser Thr Leu Ala Ala Leu Phe Met Ser Gln Val 290295 300 Phe Pro Arg Leu Lys Pro Leu Phe Val Ile Tyr Thr Leu Trp Met Trp305 310 315 320 Trp Ala Thr Met Tyr Leu Ser His His Tyr Ala Val Asp LeuVal Gly 325 330 335 Gly Gly Leu Leu Ala Thr Val Ala Phe Tyr Phe Ala LysThr Arg Phe 340 345 350 Met Pro Arg Val Gln Asn Asp Lys Met Phe Arg TrpAsp Tyr Asp Tyr 355 360 365 Val Glu Ser Gly Asp Ser Ala Leu Asp Tyr GlyTyr Gly Pro Ala Ser 370 375 380 Phe Glu Gly Glu Phe Asn Leu Asp Ser AspGlu Trp Thr Val Gly Ser 385 390 395 400 Ser Ser Ser Ile Ser Ser Gly SerLeu Ser Pro Val Asp Asp His Tyr 405 410 415 Ser Trp Glu Gly Glu Thr LeuAla Ser Pro Ala Thr Glu Ile Glu Ser 420 425 430 Gly Arg His Phe 435 152959 RNA Aspergillus fumigatus 15 auaugauuua aaauuaaugu uuccauuuucuuccccauaa caacucuucu cgcccuuccu 60 ccggcuccgu ggccaaauug uuuuaugcagcgccuccuag cgauuuaacc ucguucucgu 120 ugcccuugcc uguccgccuu gcgucaguacgacccuugca acgugaccuu ccccagagua 180 uccucguuug gccgcuggag accggagcuugcacccucau aaacuagcuc uucgaaauca 240 auucuccguu cuccagagau uaucggaucgaaucucuccg cugucgacac cuuucgucuc 300 ucggugaucc ucgcccuugg agucucgucacguugacgcc uugaaccccu ggccgccaac 360 uccacauagg agaccacacu ucauucuucccccgccauaa uugcagcacc cuccgucucc 420 cuucgagcuc cuccuggauc aucaaguccgaaaggauuag acucgucgca gcgaugaaua 480 ccacccuucc auccuggaag gaucggacgcaaaaccaguu cggcaagcuc cagauccaag 540 ucccauggcg caccauacaa cuucucgugccgcaccguau gcgacggaag auucggucca 600 agcugcgcag ucggaucucg ccuaccucaucgauauccuc guugcagacg ucauucucac 660 cugucgauac acucaggucg cugcaaagucauagauggac gcucuaugac uuucaguauc 720 uuuugcugcu gauugucggc auauucucgcugagcguuau ggaaucaccu ggaccauugg 780 caaagaccgc cgcguuuacg cuacuucucgucucucuccu ucucccgauu acgcgccagu 840 ucuucuugcc auuccucccg auugcaggauggcuuauauu uuucuacgcu ugccaguuca 900 ucccgagcga cuggcgcccu gcaaucuggguucgcgugcu gccggcucug gaaaacauuc 960 ucuacggugc uaauaucagu aacauccuuuccgcucacca aaauguggug cuugacguuu 1020 uggcguggcu ucccuacgga aucugccauuauggcgcgcc auuugugugc ucagcgauca 1080 uguucaucuu ugguccuccc ggcaccgucccccuuuucgc ucgaacuuuu ggauacauca 1140 gcauggcugc agucaccauu cagcuguuuuuccccugcuc uccuccgugg uacgaaaauc 1200 uguaugguuu ggcuccggcu gauuacuccaugccggguaa uccugcgggc cuugcucgca 1260 ucgaugagcu uuuugggaua gacuuguacacaucgggcuu cagacaaucu cccgucgugu 1320 uuggcgcauu uccuucccua caugccgcugauucgacacu ugcagcucua uuuaugagcc 1380 aaguguuccc acgguugaag cccuuguuugucaucuauac ucucuggaug uggugggcua 1440 caauguaucu uucgcaccac uacgcuguugaucuggucgg ugguggccuc uuggcaacug 1500 ucgcguucua cuuugcuaaa acgcgguucaugccucgcgu ccagaaugau aagauguucc 1560 gcugggacua cgauuauguu gagucaggcgauuccgcacu cgacuauggg uacgguccag 1620 ccagcuucga aggcgaauuc aaccuugauagcgaugagug gaccguuggu ucuucgucau 1680 ccauuucguc cggcucccuc aguccaguugacgaccacua cucuugggag ggcgagacuc 1740 uugccucucc ugccaccgag aucgagucuggaaggcauuu cugauccugc ucaaugagcc 1800 uugauacgua cuacacugug uacgugcuacugcauugacu aaugagacgg cguuuucaaa 1860 caaauuuuaa cgacaugcuu gguuaucgcauugagcugau uucgacacau auauauguuu 1920 aauacguuuu ggggacacuc cagggauucaugacgguugc uucauauccc gaccugggga 1980 uggauugacc ugguugugcc caauuuucuucugccuaacg uuuugauuau acaugcauuu 2040 uucacgaaac cagccggccc gccaugaucgugaccucaau uugagcucga aucuuccugg 2100 ggccuccagc gauaauucuu aaugcucguuccgagggugc cacaucggac auucgcuugu 2160 acaacuuuug cagaacgaac auuuucaccgauuccaacuu gagucauucg cuuacuacuu 2220 ucaacugguc gagaaacuuc gcuucuuuucagcucggcua ggugcauaaa uauuuuacau 2280 ucgugucgau cgcucacauu ucacggcgccuggaaacuug gggguuucga uuucauugga 2340 aaggauaaac aacaugggcu gggcgccuuuuacacgcacu acauucgcuu agaaaaguuc 2400 ugaugcuuuu aaugauucuu gcauuggcauauagaaaggg guccuccaga cucgcuacug 2460 ugguccucuc ucaaccccac acucgcuugcuuuuaacagu ggacaccccg uggagcuacg 2520 ucuccaucaa auauuuggca ucaaccggaaucgaugccag gaggacugag cuuacucacg 2580 gugaccgucg gguaaaaggc guucuaacagaacaucuccu cuauccuccu gucccaucuc 2640 gauucucugg cugcuggacg caacacaccucgcugcacgu uuucgacuuc cuaauacgac 2700 cuaauaucau ucucgguuuu cuuugcucuggcucgcggcc gccauuuaua uggcgugucg 2760 gcucgagucu gaagugaacu uuuuucuccuuucuggccuc cacaacuuuc cgaucccuag 2820 cagcuucccg ugcacagcga gguguuguuggaugauuguu ccauagcauu aucauuauuc 2880 cuaauccggu agcguuauga uuuaugaagaacagugaugu acauuauuau gcggugacua 2940 aaaaaaaaaa aaaaaaaaa 2959 16 2856DNA Aspergillus nidulans CDS (395)..(1714) 16 ggtttatact ccggctccgtggccatctgc ctccctcacg acctcctcgt tccaggtttt 60 cctctcgact gctgcgcccttgcacttcgc cttgcatcag tgaaaccccc tgcaacgtga 120 cggctcaaag acatcctcgtttggccgctg gagaccggag cgtgcgcttc gtttcgtctt 180 cttcgaaccg atctcaatttccccgctcgg gttgacgccg tcagcaccct gctcgttgcc 240 tcccggcttg ttattcaagaccccttttct gccgcttccg cgaccgattt attcgtcgcc 300 ttccaactct tgtacaatcggggggaaaga aagcagacgg agttcgatct ggaggaatta 360 tagctgagtc ttgcccgcaagactcgccgc aacc atg aat caa aca ctt ccc acg 415 Met Asn Gln Thr Leu ProThr 1 5 tgg aag gac cgc acg gac aac cag ttt gga aag ctt cag atc cag gtt463 Trp Lys Asp Arg Thr Asp Asn Gln Phe Gly Lys Leu Gln Ile Gln Val 1015 20 cca tgg cgg tcc atc caa ctg ctc gtc ccg cat cgc atg cgg cgg aag511 Pro Trp Arg Ser Ile Gln Leu Leu Val Pro His Arg Met Arg Arg Lys 2530 35 tta agg tcc aaa ttg cgc agt aga gcg tct cct acc tcg tca ata gcc559 Leu Arg Ser Lys Leu Arg Ser Arg Ala Ser Pro Thr Ser Ser Ile Ala 4045 50 55 tct tta cag acg tcg tta tcg cct gca gac aca cta cga tcg ctc caa607 Ser Leu Gln Thr Ser Leu Ser Pro Ala Asp Thr Leu Arg Ser Leu Gln 6065 70 agc cac cga tgg acg gtt tac gac ttc caa tat ctg ctt ctg ttg atc655 Ser His Arg Trp Thr Val Tyr Asp Phe Gln Tyr Leu Leu Leu Leu Ile 7580 85 gtg ggc atc ttc tct ttg acc gtt atc gag tcg ccc ggg cct ttg ggc703 Val Gly Ile Phe Ser Leu Thr Val Ile Glu Ser Pro Gly Pro Leu Gly 9095 100 aaa acg gcc att ttc tcc atg ctc cta ttc tct ctc ctg atc cct atg751 Lys Thr Ala Ile Phe Ser Met Leu Leu Phe Ser Leu Leu Ile Pro Met 105110 115 acc cgc cag ttc ttc ctc ccg ttt ctg ccg att gcc gga tgg ctt ctg799 Thr Arg Gln Phe Phe Leu Pro Phe Leu Pro Ile Ala Gly Trp Leu Leu 120125 130 135 ttt ttc tac gcc tgc cag ttc atc cca agc gat tgg cgc cct gcgatt 847 Phe Phe Tyr Ala Cys Gln Phe Ile Pro Ser Asp Trp Arg Pro Ala Ile140 145 150 tgg gtt cgt gtc ttg cct gca ctg gag aat att ctc tac ggc gcaaac 895 Trp Val Arg Val Leu Pro Ala Leu Glu Asn Ile Leu Tyr Gly Ala Asn155 160 165 atc agc aac atc cta tcc gct cac cag aac gtt gtg ctt gac gtgctg 943 Ile Ser Asn Ile Leu Ser Ala His Gln Asn Val Val Leu Asp Val Leu170 175 180 gcg tgg cta ccc tac ggt atc tgc cac tat ggc gct ccg ttt gtgtgc 991 Ala Trp Leu Pro Tyr Gly Ile Cys His Tyr Gly Ala Pro Phe Val Cys185 190 195 tcg ttg atc atg ttc atc ttc ggt ccg ccc ggc act gtt ccc cttttc 1039 Ser Leu Ile Met Phe Ile Phe Gly Pro Pro Gly Thr Val Pro Leu Phe200 205 210 215 gcg cgc act ttc ggc tat atc agt atg act gcg gtt act attcag ctg 1087 Ala Arg Thr Phe Gly Tyr Ile Ser Met Thr Ala Val Thr Ile GlnLeu 220 225 230 ttt ttc cct tgc tct cca cct tgg tat gag aat cgc tat ggtcta gct 1135 Phe Phe Pro Cys Ser Pro Pro Trp Tyr Glu Asn Arg Tyr Gly LeuAla 235 240 245 ccg gca gac tac tcc atc caa ggt gat ccc gca ggg ctt gcccgc att 1183 Pro Ala Asp Tyr Ser Ile Gln Gly Asp Pro Ala Gly Leu Ala ArgIle 250 255 260 gac aag ctt ttc ggc atc gac ctt tac acg tct gtt ttc catcag tcg 1231 Asp Lys Leu Phe Gly Ile Asp Leu Tyr Thr Ser Val Phe His GlnSer 265 270 275 cct gtt gtg ttc ggc gct ttt ccg tcg ctg cat gct gcc gactca acc 1279 Pro Val Val Phe Gly Ala Phe Pro Ser Leu His Ala Ala Asp SerThr 280 285 290 295 ctg gcc gca ctt ttc atg agt cat gtt ttc ccc cgc atgaag ccc gtc 1327 Leu Ala Ala Leu Phe Met Ser His Val Phe Pro Arg Met LysPro Val 300 305 310 ttc gtg acc tat act cta tgg atg tgg tgg gca aca atgtac ctc tca 1375 Phe Val Thr Tyr Thr Leu Trp Met Trp Trp Ala Thr Met TyrLeu Ser 315 320 325 cat cac tat gcg gtc gat ttg gtt gcg ggt ggt ctc ctggcc gcc att 1423 His His Tyr Ala Val Asp Leu Val Ala Gly Gly Leu Leu AlaAla Ile 330 335 340 gct ttc tac ttc gcc aag acc cga ttc ctt ccc cgt gtccag ctc gac 1471 Ala Phe Tyr Phe Ala Lys Thr Arg Phe Leu Pro Arg Val GlnLeu Asp 345 350 355 aag acc ttc cgt tgg gac tac gac tat gtg gaa ttc ggcgag tct gcc 1519 Lys Thr Phe Arg Trp Asp Tyr Asp Tyr Val Glu Phe Gly GluSer Ala 360 365 370 375 ctg gag tat ggg tat ggt gca gct ggc tat gat ggagac ttc aat ctc 1567 Leu Glu Tyr Gly Tyr Gly Ala Ala Gly Tyr Asp Gly AspPhe Asn Leu 380 385 390 gac agc gat gaa tgg act gtt ggt tct tca tcc tccgtc tcc tca ggc 1615 Asp Ser Asp Glu Trp Thr Val Gly Ser Ser Ser Ser ValSer Ser Gly 395 400 405 tcc ttg agt ccc gtt gac gat cat tac tca tgg gaaacc gag gca ctg 1663 Ser Leu Ser Pro Val Asp Asp His Tyr Ser Trp Glu ThrGlu Ala Leu 410 415 420 acc tcc cca cat act gat att gag tcc ggc agg catact ttc agc cct 1711 Thr Ser Pro His Thr Asp Ile Glu Ser Gly Arg His ThrPhe Ser Pro 425 430 435 tga gtagccacaa accaaactcg atacctgcat atagcgatctcgctcctcct 1764 ccactgcatc tatttacgag acggcgttag aacatttcac gacattctggctttattgca 1824 tcgagcacat ttcgacacat atatctttaa taccctttct tcggtgtcccagatcatcgg 1884 ttcgacctta atgtacctcg gtccgaatcc gcctgggata ctgtttctctttccgccgca 1944 cttcactgta cattgcttga cattgcgaaa ccgggttggg ctcgaacgtgggatgggtta 2004 tcgctcatcg ctacacgccg ttgctccatc ataatgttaa tggacacaatggggctacgc 2064 atcctggtgt ttagtcctgg aagaccatcc gataaccccc gtcggtaacactcgcttgtc 2124 tcgtgtccac ccagacacta cttcaattct cacttctatc gtccgctattaccttgacct 2184 ggtcgaaccc atccttatta ttcgtttcga ctatgctata tatttatttttaccattcgt 2244 gtcgatcgct catactcttg gcgcttggga ctggaagcat ttatattggaaaaaatcacg 2304 gaatggggcg ccttttcttc ttgcacttca ctcgctgtgc atagacggttttacatttct 2364 gctttgcaat gcatcacgaa ctctgcatta gcatatagaa agaggggaaggatggacctt 2424 cttcttgatt gctcgcatgg tttatccatt cgctcaaagt ggattacgtccacatcttac 2484 ccgggggcta tacacatggc tactgtgttg ctttctgaca ttcgccggacgtgcaaggtt 2544 gggaggagag tctgacgctg acggggcttg ttgaaggatg ttcacgcgtcccgatttgac 2604 ccggcttcga ctaacctcag attctcgact tgttggacgg tgacttgacttgcttgctat 2664 ggtctgacgc tctcacacct acctatcaca tcctcctcac ctcacaaattccgctcatgg 2724 acactatcct cttcttttcg tttcccttgg atagtgtgtg tgtgtgtgtggttggggcaa 2784 attatccata gcagcagtat tattagttat aatccggtag tgttatgatttatgaaggca 2844 acttgtatac ta 2856 17 439 PRT Aspergillus nidulans 17Met Asn Gln Thr Leu Pro Thr Trp Lys Asp Arg Thr Asp Asn Gln Phe 1 5 1015 Gly Lys Leu Gln Ile Gln Val Pro Trp Arg Ser Ile Gln Leu Leu Val 20 2530 Pro His Arg Met Arg Arg Lys Leu Arg Ser Lys Leu Arg Ser Arg Ala 35 4045 Ser Pro Thr Ser Ser Ile Ala Ser Leu Gln Thr Ser Leu Ser Pro Ala 50 5560 Asp Thr Leu Arg Ser Leu Gln Ser His Arg Trp Thr Val Tyr Asp Phe 65 7075 80 Gln Tyr Leu Leu Leu Leu Ile Val Gly Ile Phe Ser Leu Thr Val Ile 8590 95 Glu Ser Pro Gly Pro Leu Gly Lys Thr Ala Ile Phe Ser Met Leu Leu100 105 110 Phe Ser Leu Leu Ile Pro Met Thr Arg Gln Phe Phe Leu Pro PheLeu 115 120 125 Pro Ile Ala Gly Trp Leu Leu Phe Phe Tyr Ala Cys Gln PheIle Pro 130 135 140 Ser Asp Trp Arg Pro Ala Ile Trp Val Arg Val Leu ProAla Leu Glu 145 150 155 160 Asn Ile Leu Tyr Gly Ala Asn Ile Ser Asn IleLeu Ser Ala His Gln 165 170 175 Asn Val Val Leu Asp Val Leu Ala Trp LeuPro Tyr Gly Ile Cys His 180 185 190 Tyr Gly Ala Pro Phe Val Cys Ser LeuIle Met Phe Ile Phe Gly Pro 195 200 205 Pro Gly Thr Val Pro Leu Phe AlaArg Thr Phe Gly Tyr Ile Ser Met 210 215 220 Thr Ala Val Thr Ile Gln LeuPhe Phe Pro Cys Ser Pro Pro Trp Tyr 225 230 235 240 Glu Asn Arg Tyr GlyLeu Ala Pro Ala Asp Tyr Ser Ile Gln Gly Asp 245 250 255 Pro Ala Gly LeuAla Arg Ile Asp Lys Leu Phe Gly Ile Asp Leu Tyr 260 265 270 Thr Ser ValPhe His Gln Ser Pro Val Val Phe Gly Ala Phe Pro Ser 275 280 285 Leu HisAla Ala Asp Ser Thr Leu Ala Ala Leu Phe Met Ser His Val 290 295 300 PhePro Arg Met Lys Pro Val Phe Val Thr Tyr Thr Leu Trp Met Trp 305 310 315320 Trp Ala Thr Met Tyr Leu Ser His His Tyr Ala Val Asp Leu Val Ala 325330 335 Gly Gly Leu Leu Ala Ala Ile Ala Phe Tyr Phe Ala Lys Thr Arg Phe340 345 350 Leu Pro Arg Val Gln Leu Asp Lys Thr Phe Arg Trp Asp Tyr AspTyr 355 360 365 Val Glu Phe Gly Glu Ser Ala Leu Glu Tyr Gly Tyr Gly AlaAla Gly 370 375 380 Tyr Asp Gly Asp Phe Asn Leu Asp Ser Asp Glu Trp ThrVal Gly Ser 385 390 395 400 Ser Ser Ser Val Ser Ser Gly Ser Leu Ser ProVal Asp Asp His Tyr 405 410 415 Ser Trp Glu Thr Glu Ala Leu Thr Ser ProHis Thr Asp Ile Glu Ser 420 425 430 Gly Arg His Thr Phe Ser Pro 435 182877 RNA Aspergillus nidulans 18 uaaaauuuca uaaauguuuc cgguuuauacuccggcuccg uggccaucug ccucccucac 60 gaccuccucg uuccagguuu uccucucgacugcugcgccc uugcacuucg ccuugcauca 120 gugaaacccc cugcaacgug acggcucaaagacauccucg uuuggccgcu ggagaccgga 180 gcgugcgcuu cguuucgucu ucuucgaaccgaucucaauu uccccgcucg gguugacgcc 240 gucagcaccc ugcucguugc cucccggcuuguuauucaag accccuuuuc ugccgcuucc 300 gcgaccgauu uauucgucgc cuuccaacucuuguacaauc ggggggaaag aaagcagacg 360 gaguucgauc uggaggaauu auagcugagucuugcccgca agacucgccg caaccaugaa 420 ucaaacacuu cccacgugga aggaccgcacggacaaccag uuuggaaagc uucagaucca 480 gguuccaugg cgguccaucc aacugcucgucccgcaucgc augcggcgga aguuaagguc 540 caaauugcgc aguagagcgu cuccuaccucgucaauagcc ucuuuacaga cgucguuauc 600 gccugcagac acacuacgau cgcuccaaagccaccgaugg acgguuuacg acuuccaaua 660 ucugcuucug uugaucgugg gcaucuucucuuugaccguu aucgagucgc ccgggccuuu 720 gggcaaaacg gccauuuucu ccaugcuccuauucucucuc cugaucccua ugacccgcca 780 guucuuccuc ccguuucugc cgauugccggauggcuucug uuuuucuacg ccugccaguu 840 caucccaagc gauuggcgcc cugcgauuuggguucguguc uugccugcac uggagaauau 900 ucucuacggc gcaaacauca gcaacauccuauccgcucac cagaacguug ugcuugacgu 960 gcuggcgugg cuacccuacg guaucugccacuauggcgcu ccguuugugu gcucguugau 1020 cauguucauc uucgguccgc ccggcacuguuccccuuuuc gcgcgcacuu ucggcuauau 1080 caguaugacu gcgguuacua uucagcuguuuuucccuugc ucuccaccuu gguaugagaa 1140 ucgcuauggu cuagcuccgg cagacuacuccauccaaggu gaucccgcag ggcuugcccg 1200 cauugacaag cuuuucggca ucgaccuuuacacgucuguu uuccaucagu cgccuguugu 1260 guucggcgcu uuuccgucgc ugcaugcugccgacucaacc cuggccgcac uuuucaugag 1320 ucauguuuuc ccccgcauga agcccgucuucgugaccuau acucuaugga uguggugggc 1380 aacaauguac cucucacauc acuaugcggucgauuugguu gcgggugguc uccuggccgc 1440 cauugcuuuc uacuucgcca agacccgauuccuuccccgu guccagcucg acaagaccuu 1500 ccguugggac uacgacuaug uggaauucggcgagucugcc cuggaguaug gguauggugc 1560 agcuggcuau gauggagacu ucaaucucgacagcgaugaa uggacuguug guucuucauc 1620 cuccgucucc ucaggcuccu ugagucccguugacgaucau uacucauggg aaaccgaggc 1680 acugaccucc ccacauacug auauugaguccggcaggcau acuuucagcc cuugaguagc 1740 cacaaaccaa acucgauacc ugcauauagcgaucucgcuc cuccuccacu gcaucuauuu 1800 acgagacggc guuagaacau uucacgacauucuggcuuua uugcaucgag cacauuucga 1860 cacauauauc uuuaauaccc uuucuucggugucccagauc aucgguucga ccuuaaugua 1920 ccucgguccg aauccgccug ggauacuguuucucuuuccg ccgcacuuca cuguacauug 1980 cuugacauug cgaaaccggg uugggcucgaacgugggaug gguuaucgcu caucgcuaca 2040 cgccguugcu ccaucauaau guuaauggacacaauggggc uacgcauccu gguguuuagu 2100 ccuggaagac cauccgauaa cccccgucgguaacacucgc uugucucgug uccacccaga 2160 cacuacuuca auucucacuu cuaucguccgcuauuaccuu gaccuggucg aacccauccu 2220 uauuauucgu uucgacuaug cuauauauuuauuuuuacca uucgugucga ucgcucauac 2280 ucuuggcgcu ugggacugga agcauuuauauuggaaaaaa ucacggaaug gggcgccuuu 2340 ucuucuugca cuucacucgc ugugcauagacgguuuuaca uuucugcuuu gcaaugcauc 2400 acgaacucug cauuagcaua uagaaagaggggaaggaugg accuucuucu ugauugcucg 2460 caugguuuau ccauucgcuc aaaguggauuacguccacau cuuacccggg ggcuauacac 2520 auggcuacug uguugcuuuc ugacauucgccggacgugca agguugggag gagagucuga 2580 cgcugacggg gcuuguugaa ggauguucacgcgucccgau uugacccggc uucgacuaac 2640 cucagauucu cgacuuguug gacggugacuugacuugcuu gcuauggucu gacgcucuca 2700 caccuaccua ucacauccuc cucaccucacaaauuccgcu cauggacacu auccucuucu 2760 uuucguuucc cuuggauagu gugugugugugugugguugg ggcaaauuau ccauagcagc 2820 aguauuauua guuauaaucc gguaguguuaugauuuauga aggcaacuug uauacua 2877 19 3220 DNA Candida neoformans intron(1888)..(1939) 19 gtcgacatag ttacactccc ctctactctt cagactaccc cttctgattatcaggagact 60 acgactactc cgcctacctt gacctttgca aactcggagc cacgaccgagtccttccctc 120 catcgttccc tgcgcaacat cggtttggcc gacggcagca tgacttgtgccccaggctat 180 atcctcatct gcattcatca tgaccccttg tctcgttaca cgcttggcatcaattttacc 240 acctttgtgg gtctttgccc cacgacccct gctagtacca gcacttccggcggtcccgcc 300 ttttcctctt ttacacccaa gttcgccgac catatgcgat gtcatgtcatccagcgcttg 360 ctgcacactg tcatgcacga ccgtctcgcc aacatgccgg gttgccacacctgggaatcg 420 ttggtcaatg cggcgtaccg ttactgcacc tgggctatgg aatgtatcttcctcggcacc 480 gatgcaaatg ttcaattcgg gattgcttgt tatccccact tgaaagagctcgaagaggcg 540 gcaaggacgc tcaggaagga tagtatgggc gagtgggatg atgtggtgcagattggggtg 600 gataggttgt tggaaatact accttgtggt gaacggacct tgatcaacaagagaaggatc 660 gaaaaggaac gactgcaagc gcttctaaaa gctcaaactc aagtcaaggccaagcatgag 720 gccagatgtc aggtggcaac cccgatccca gctccagctg tcattcctgtctctgcacct 780 gctccggtta taatcaagcg taaaaaatac agtagcctac gcaagctgatgggacgtgcg 840 gggtcggtga ttggcaggca gaagatgcgg gcaagggcgt ctgcgattatcgagaaggga 900 aaggccggct tctacgatag gcaaaaggtc cagcgcatgg agccggctccatccatgtct 960 cgtcctgcgg gagtgtcttg tgtgtgattg tcagctggcc gtgaagatagtcttggagct 1020 gtgtataggg atactacatg atgtgcatgc agtgtgtttt caaagcaattgccacgtggt 1080 gttcgcgtgg gtcggaaacg aggttcgcgt tgttatggtt cttgttttgagattctcgcg 1140 ctggctacac gtacgcacca tacactgcag acgacagcgg caagcatacagacggcaacg 1200 ctgacatccc agccacacac accatgtccg ccatccgcgc actcacgagccccgtcaccg 1260 cctgcttctc cacctctctc tccccgcatg ccgcattcca ccgcttcctccatgcgctct 1320 ccgcctccat ccgccgcctc gacctctccc gcgacccgcg caagaccttggaccgcctca 1380 ggcagcacag gttcaccctt gccaacactt tgccccgcgc gttcatgcttctgtgcgcct 1440 cctacagtct ctacatcatg accaccccgc ccttcccact caagctcgggatacccatcg 1500 catacatcgc cgctgtcatc ttcccgatca cgtcgcagtt tgtctggccggcgacaccca 1560 tctttgcatg gctcatcacc tttttctccg cccgtttcat cccttccggccgccggcccg 1620 aaatccacgt cgcccttctc ccagcccttg aatccgtcct ctacggtgccaacatttctg 1680 atctccaaac acgatatacc aacgctttcc ttgatgtggt cgcctggctgccctatgggg 1740 tattacattt cacccttcca ttcgtcgtgg ctgttatcct ttggtcactgggacctaggg 1800 gtgcggttca gttctggggt ttggcctttg ggtggatgaa tttgctcggtgtcgtctgcc 1860 agctcctctt ccctgctgcc gcgccttgta agtacctctc tcttttatttaatatatcca 1920 tacttaatat gccgggcagg gtacgaaatc attcacggtc ttacccccgccgactattcc 1980 atggctggct ctcccggcgg tctcatgcgt atcgaccgcg tcttccattcttcaggctat 2040 accaacgcat tcggttccgc tcctcttgtt tttggcgcct tcccctccctccattcagga 2100 tgtgccgtca tggaagccct cttcctctcc catttcttcc catccctcaagggcctgtat 2160 tggggctatg tcggtgtcct ctggtgggcg accatgtacc tctcccaccactacctcatc 2220 gacctcgttg gcggagcttg tctgagcgtc ttggtgttct acttgtgcatgcctgaaggt 2280 ttcaaggacg ttgatcagat acagtgggag gcggtggaag gggatggatacgagatgatt 2340 ggcggaccca gaacaggcac aggtcctgag attgatttgg atgaggaaattagaaagttg 2400 gaagaacaag gcgaagcgct ttttgagcag gtgatagggg atgaagagtctcggatcgag 2460 acaaggggag aaggaaacag tggcgcgggc gggaatctga gtgggaatgaaagtggtgat 2520 agtgcagata gcggtagggg aaagggcaag gggaagggga aggggaaacaaactgccaaa 2580 aagccaaagg cgaaggagca aagatcggtt agctggggag aaacaaaggtgatgggtgaa 2640 ggagcgcagg tggcttcaga gaatagctct tagatatctg ttttaaaacagtagtatacc 2700 caacaatttt ttgacgatat accctaccac tatgatctta ataatgcagaatagaaaaac 2760 accttgcccc tctcgcaggc gtcaacacat ggagtctagg cagttaaatcacactacatc 2820 atcatcgccg aaaaaaataa acctagtaac tccaacccaa ccctcctggtacaggcctgt 2880 tcgtccaccg actgataatt cccatcgccc tcgccctcct cacaagtttacccacccgcc 2940 tttgcgactt ccaagtcaac ccggtctccg cccgacttct tatcttgcccatcgggttga 3000 cgaattcata ggcgaaatta gggttcatat cgtgcaccaa ggggttcgttttggtgatgt 3060 aaaaaggatc aagcttgacg gcgattttct tgggaggacc aagaagaggtggacggggca 3120 aaggtcgagc ggtggggtag atggattctt gggtgaatgc ttgaggggcagtaaactaga 3180 agagttgaga aaacccaaag gagtcagcct ttatgtcgac 3220 20 3120DNA Cryptococcus neoformans CDS (1224)..(2624) 20 gtcgacatag ttacactcccctctactctt cagactaccc cttctgatta tcaggagact 60 acgactactc cgcctaccttgacctttgca aactcggagc cacgaccgag tccttccctc 120 catcgttccc tgcgcaacatcggtttggcc gacggcagca tgacttgtgc cccaggctat 180 atcctcatct gcattcatcatgaccccttg tctcgttaca cgcttggcat caattttacc 240 acctttgtgg gtctttgccccacgacccct gctagtacca gcacttccgg cggtcccgcc 300 ttttcctctt ttacacccaagttcgccgac catatgcgat gtcatgtcat ccagcgcttg 360 ctgcacactg tcatgcacgaccgtctcgcc aacatgccgg gttgccacac ctgggaatcg 420 ttggtcaatg cggcgtaccgttactgcacc tgggctatgg aatgtatctt cctcggcacc 480 gatgcaaatg ttcaattcgggattgcttgt tatccccact tgaaagagct cgaagaggcg 540 gcaaggacgc tcaggaaggatagtatgggc gagtgggatg atgtggtgca gattggggtg 600 gataggttgt tggaaatactaccttgtggt gaacggacct tgatcaacaa gagaaggatc 660 gaaaaggaac gactgcaagcgcttctaaaa gctcaaactc aagtcaaggc caagcatgag 720 gccagatgtc aggtggcaaccccgatccca gctccagctg tcattcctgt ctctgcacct 780 gctccggtta taatcaagcgtaaaaaatac agtagcctac gcaagctgat gggacgtgcg 840 gggtcggtga ttggcaggcagaagatgcgg gcaagggcgt ctgcgattat cgagaaggga 900 aaggccggct tctacgataggcaaaaggtc cagcgcatgg agccggctcc atccatgtct 960 cgtcctgcgg gagtgtcttgtgtgtgattg tcagctggcc gtgaagatag tcttggagct 1020 gtgtataggg atactacatgatgtgcatgc agtgtgtttt caaagcaatt gccacgtggt 1080 gttcgcgtgg gtcggaaacgaggttcgcgt tgttatggtt cttgttttga gattctcgcg 1140 ctggctacac gtacgcaccatacactgcag acgacagcgg caagcataca gacggcaacg 1200 ctgacatccc agccacacacacc atg tcc gcc atc cgc gca ctc acg agc ccc 1253 Met Ser Ala Ile Arg AlaLeu Thr Ser Pro 1 5 10 gtc acc gcc tgc ttc tcc acc tct ctc tcc ccg catgcc gca ttc cac 1301 Val Thr Ala Cys Phe Ser Thr Ser Leu Ser Pro His AlaAla Phe His 15 20 25 cgc ttc ctc cat gcg ctc tcc gcc tcc atc cgc cgc ctcgac ctc tcc 1349 Arg Phe Leu His Ala Leu Ser Ala Ser Ile Arg Arg Leu AspLeu Ser 30 35 40 cgc gac ccg cgc aag acc ttg gac cgc ctc agg cag cac aggttc acc 1397 Arg Asp Pro Arg Lys Thr Leu Asp Arg Leu Arg Gln His Arg PheThr 45 50 55 ctt gcc aac act ttg ccc cgc gcg ttc atg ctt ctg tgc gcc tcctac 1445 Leu Ala Asn Thr Leu Pro Arg Ala Phe Met Leu Leu Cys Ala Ser Tyr60 65 70 agt ctc tac atc atg acc acc ccg ccc ttc cca ctc aag ctc ggg ata1493 Ser Leu Tyr Ile Met Thr Thr Pro Pro Phe Pro Leu Lys Leu Gly Ile 7580 85 90 ccc atc gca tac atc gcc gct gtc atc ttc ccg atc acg tcg cag tcc1541 Pro Ile Ala Tyr Ile Ala Ala Val Ile Phe Pro Ile Thr Ser Gln Ser 95100 105 gcc cgt ttc atc cct tcc ggc cgc cgg ccc gaa atc cac gtc gcc ctt1589 Ala Arg Phe Ile Pro Ser Gly Arg Arg Pro Glu Ile His Val Ala Leu 110115 120 ctc cca gcc ctt gaa tcc gtc ctc tac ggt gcc aac att tct gat ctc1637 Leu Pro Ala Leu Glu Ser Val Leu Tyr Gly Ala Asn Ile Ser Asp Leu 125130 135 caa aca cga tat acc aac gct ttc ctt gat gtg gtc gcc tgg ctg ccc1685 Gln Thr Arg Tyr Thr Asn Ala Phe Leu Asp Val Val Ala Trp Leu Pro 140145 150 tat ggg gta tta cat ttc acc ctt cca ttc gtc gtg gct gtt atc ctt1733 Tyr Gly Val Leu His Phe Thr Leu Pro Phe Val Val Ala Val Ile Leu 155160 165 170 tgg tca ctg gga cct agg ggt gcg gtt cag ttc tgg ggt ttg gccttt 1781 Trp Ser Leu Gly Pro Arg Gly Ala Val Gln Phe Trp Gly Leu Ala Phe175 180 185 ggg tgg atg aat ttg ctc ggt gtc gtc tgc cag ctc ctc ttc cctgct 1829 Gly Trp Met Asn Leu Leu Gly Val Val Cys Gln Leu Leu Phe Pro Ala190 195 200 gcc gcg cct tgg tac gaa atc att cac ggt ctt acc ccc gcc gactat 1877 Ala Ala Pro Trp Tyr Glu Ile Ile His Gly Leu Thr Pro Ala Asp Tyr205 210 215 tcc atg gct ggc tct ccc ggc ggt ctc atg cgt atc gac cgc gtcttc 1925 Ser Met Ala Gly Ser Pro Gly Gly Leu Met Arg Ile Asp Arg Val Phe220 225 230 cat tct tca ggc tat acc aac gca ttc ggt tcc gct cct ctt gttttt 1973 His Ser Ser Gly Tyr Thr Asn Ala Phe Gly Ser Ala Pro Leu Val Phe235 240 245 250 ggc gcc ttc ccc tcc ctc cat tca gga tgt gcc gtc atg gaagcc ctc 2021 Gly Ala Phe Pro Ser Leu His Ser Gly Cys Ala Val Met Glu AlaLeu 255 260 265 ttc ctc tcc cat ttc ttc cca tcc ctc aag ggc ctg tat tggggc tat 2069 Phe Leu Ser His Phe Phe Pro Ser Leu Lys Gly Leu Tyr Trp GlyTyr 270 275 280 gtc ggt gtc ctc tgg tgg gcg acc atg tac ctc tcc cac cactac ctc 2117 Val Gly Val Leu Trp Trp Ala Thr Met Tyr Leu Ser His His TyrLeu 285 290 295 atc gac ctc gtt ggc gga gct tgt ctg agc gtc ttg gtg ttctac ttg 2165 Ile Asp Leu Val Gly Gly Ala Cys Leu Ser Val Leu Val Phe TyrLeu 300 305 310 tgc atg cct gaa ggt ttc aag gac gtt gat cag ata cag tgggag gcg 2213 Cys Met Pro Glu Gly Phe Lys Asp Val Asp Gln Ile Gln Trp GluAla 315 320 325 330 gtg gaa ggg gat gga tac gag atg att ggc gga ccc agaaca ggc aca 2261 Val Glu Gly Asp Gly Tyr Glu Met Ile Gly Gly Pro Arg ThrGly Thr 335 340 345 ggt cct gag att gat ttg gat gag gaa att aga aag ttggaa gaa caa 2309 Gly Pro Glu Ile Asp Leu Asp Glu Glu Ile Arg Lys Leu GluGlu Gln 350 355 360 ggc gaa gcg ctt ttt gag cag gtg ata ggg gat gaa gagtct cgg atc 2357 Gly Glu Ala Leu Phe Glu Gln Val Ile Gly Asp Glu Glu SerArg Ile 365 370 375 gag aca agg gga gaa gga aac agt ggc gcg ggc ggg aatctg agt ggg 2405 Glu Thr Arg Gly Glu Gly Asn Ser Gly Ala Gly Gly Asn LeuSer Gly 380 385 390 aat gaa agt ggt gat agt gca gat agc ggt agg gga aagggc aag ggg 2453 Asn Glu Ser Gly Asp Ser Ala Asp Ser Gly Arg Gly Lys GlyLys Gly 395 400 405 410 aag ggg aag ggg aaa caa act gcc aaa aag cca aaggcg aag gag caa 2501 Lys Gly Lys Gly Lys Gln Thr Ala Lys Lys Pro Lys AlaLys Glu Gln 415 420 425 aga tcg gtt agc tgg gga gaa aca aag gtg atg ggtgaa gga gcg cag 2549 Arg Ser Val Ser Trp Gly Glu Thr Lys Val Met Gly GluGly Ala Gln 430 435 440 gtg gct tca gag aat agc tct tag ata tct gtt ttaaaa cag tag tat 2597 Val Ala Ser Glu Asn Ser Ser Ile Ser Val Leu Lys GlnTyr 445 450 455 acc caa caa ttt ttt gac gat ata ccc taccactatgatcttaataa 2644 Thr Gln Gln Phe Phe Asp Asp Ile Pro 460 465 tgcagaatagaaaaacacct tgcccctctc gcaggcgtca acacatggag tctaggcagt 2704 taaatcacactacatcatca tcgccgaaaa aaataaacct agtaactcca acccaaccct 2764 cctggtacaggcctgttcgt ccaccgactg ataattccca tcgccctcgc cctcctcaca 2824 agtttacccacccgcctttg cgacttccaa gtcaacccgg tctccgcccg acttcttatc 2884 ttgcccatcgggttgacgaa ttcataggcg aaattagggt tcatatcgtg caccaagggg 2944 ttcgttttggtgatgtaaaa aggatcaagc ttgacggcga ttttcttggg aggaccaaga 3004 agaggtggacggggcaaagg tcgagcggtg gggtagatgg attcttgggt gaatgcttga 3064 ggggcagtaaactagaagag ttgagaaaac ccaaaggagt cagcctttat gtcgac 3120 21 449 PRTCryptococcus neoformans 21 Met Ser Ala Ile Arg Ala Leu Thr Ser Pro ValThr Ala Cys Phe Ser 1 5 10 15 Thr Ser Leu Ser Pro His Ala Ala Phe HisArg Phe Leu His Ala Leu 20 25 30 Ser Ala Ser Ile Arg Arg Leu Asp Leu SerArg Asp Pro Arg Lys Thr 35 40 45 Leu Asp Arg Leu Arg Gln His Arg Phe ThrLeu Ala Asn Thr Leu Pro 50 55 60 Arg Ala Phe Met Leu Leu Cys Ala Ser TyrSer Leu Tyr Ile Met Thr 65 70 75 80 Thr Pro Pro Phe Pro Leu Lys Leu GlyIle Pro Ile Ala Tyr Ile Ala 85 90 95 Ala Val Ile Phe Pro Ile Thr Ser GlnSer Ala Arg Phe Ile Pro Ser 100 105 110 Gly Arg Arg Pro Glu Ile His ValAla Leu Leu Pro Ala Leu Glu Ser 115 120 125 Val Leu Tyr Gly Ala Asn IleSer Asp Leu Gln Thr Arg Tyr Thr Asn 130 135 140 Ala Phe Leu Asp Val ValAla Trp Leu Pro Tyr Gly Val Leu His Phe 145 150 155 160 Thr Leu Pro PheVal Val Ala Val Ile Leu Trp Ser Leu Gly Pro Arg 165 170 175 Gly Ala ValGln Phe Trp Gly Leu Ala Phe Gly Trp Met Asn Leu Leu 180 185 190 Gly ValVal Cys Gln Leu Leu Phe Pro Ala Ala Ala Pro Trp Tyr Glu 195 200 205 IleIle His Gly Leu Thr Pro Ala Asp Tyr Ser Met Ala Gly Ser Pro 210 215 220Gly Gly Leu Met Arg Ile Asp Arg Val Phe His Ser Ser Gly Tyr Thr 225 230235 240 Asn Ala Phe Gly Ser Ala Pro Leu Val Phe Gly Ala Phe Pro Ser Leu245 250 255 His Ser Gly Cys Ala Val Met Glu Ala Leu Phe Leu Ser His PhePhe 260 265 270 Pro Ser Leu Lys Gly Leu Tyr Trp Gly Tyr Val Gly Val LeuTrp Trp 275 280 285 Ala Thr Met Tyr Leu Ser His His Tyr Leu Ile Asp LeuVal Gly Gly 290 295 300 Ala Cys Leu Ser Val Leu Val Phe Tyr Leu Cys MetPro Glu Gly Phe 305 310 315 320 Lys Asp Val Asp Gln Ile Gln Trp Glu AlaVal Glu Gly Asp Gly Tyr 325 330 335 Glu Met Ile Gly Gly Pro Arg Thr GlyThr Gly Pro Glu Ile Asp Leu 340 345 350 Asp Glu Glu Ile Arg Lys Leu GluGlu Gln Gly Glu Ala Leu Phe Glu 355 360 365 Gln Val Ile Gly Asp Glu GluSer Arg Ile Glu Thr Arg Gly Glu Gly 370 375 380 Asn Ser Gly Ala Gly GlyAsn Leu Ser Gly Asn Glu Ser Gly Asp Ser 385 390 395 400 Ala Asp Ser GlyArg Gly Lys Gly Lys Gly Lys Gly Lys Gly Lys Gln 405 410 415 Thr Ala LysLys Pro Lys Ala Lys Glu Gln Arg Ser Val Ser Trp Gly 420 425 430 Glu ThrLys Val Met Gly Glu Gly Ala Gln Val Ala Ser Glu Asn Ser 435 440 445 Ser22 6 PRT Cryptococcus neoformans 22 Ile Ser Val Leu Lys Gln 1 5 23 10PRT Cryptococcus neoformans 23 Tyr Thr Gln Gln Phe Phe Asp Asp Ile Pro 15 10 24 3168 RNA Candida neoformans 24 gucgacauag uuacacuccc cucuacucuucagacuaccc cuucugauua ucaggagacu 60 acgacuacuc cgccuaccuu gaccuuugcaaacucggagc cacgaccgag uccuucccuc 120 caucguuccc ugcgcaacau cgguuuggccgacggcagca ugacuugugc cccaggcuau 180 auccucaucu gcauucauca ugaccccuugucucguuaca cgcuuggcau caauuuuacc 240 accuuugugg gucuuugccc cacgaccccugcuaguacca gcacuuccgg cggucccgcc 300 uuuuccucuu uuacacccaa guucgccgaccauaugcgau gucaugucau ccagcgcuug 360 cugcacacug ucaugcacga ccgucucgccaacaugccgg guugccacac cugggaaucg 420 uuggucaaug cggcguaccg uuacugcaccugggcuaugg aauguaucuu ccucggcacc 480 gaugcaaaug uucaauucgg gauugcuuguuauccccacu ugaaagagcu cgaagaggcg 540 gcaaggacgc ucaggaagga uaguaugggcgagugggaug auguggugca gauuggggug 600 gauagguugu uggaaauacu accuuguggugaacggaccu ugaucaacaa gagaaggauc 660 gaaaaggaac gacugcaagc gcuucuaaaagcucaaacuc aagucaaggc caagcaugag 720 gccagauguc agguggcaac cccgaucccagcuccagcug ucauuccugu cucugcaccu 780 gcuccgguua uaaucaagcg uaaaaaauacaguagccuac gcaagcugau gggacgugcg 840 gggucgguga uuggcaggca gaagaugcgggcaagggcgu cugcgauuau cgagaaggga 900 aaggccggcu ucuacgauag gcaaaagguccagcgcaugg agccggcucc auccaugucu 960 cguccugcgg gagugucuug ugugugauugucagcuggcc gugaagauag ucuuggagcu 1020 guguauaggg auacuacaug augugcaugcaguguguuuu caaagcaauu gccacguggu 1080 guucgcgugg gucggaaacg agguucgcguuguuaugguu cuuguuuuga gauucucgcg 1140 cuggcuacac guacgcacca uacacugcagacgacagcgg caagcauaca gacggcaacg 1200 cugacauccc agccacacac accauguccgccauccgcgc acucacgagc cccgucaccg 1260 ccugcuucuc caccucucuc uccccgcaugccgcauucca ccgcuuccuc caugcgcucu 1320 ccgccuccau ccgccgccuc gaccucucccgcgacccgcg caagaccuug gaccgccuca 1380 ggcagcacag guucacccuu gccaacacuuugccccgcgc guucaugcuu cugugcgccu 1440 ccuacagucu cuacaucaug accaccccgcccuucccacu caagcucggg auacccaucg 1500 cauacaucgc cgcugucauc uucccgaucacgucgcaguu ugucuggccg gcgacaccca 1560 ucuuugcaug gcucaucacc uuuuucuccgcccguuucau cccuuccggc cgccggcccg 1620 aaauccacgu cgcccuucuc ccagcccuugaauccguccu cuacggugcc aacauuucug 1680 aucuccaaac acgauauacc aacgcuuuccuugauguggu cgccuggcug cccuaugggg 1740 uauuacauuu cacccuucca uucgucguggcuguuauccu uuggucacug ggaccuaggg 1800 gugcgguuca guucuggggu uuggccuuuggguggaugaa uuugcucggu gucgucugcc 1860 agcuccucuu cccugcugcc gcgccuugguacgaaaucau ucacggucuu acccccgccg 1920 acuauuccau ggcuggcucu cccggcggucucaugcguau cgaccgcguc uuccauucuu 1980 caggcuauac caacgcauuc gguuccgcuccucuuguuuu uggcgccuuc cccucccucc 2040 auucaggaug ugccgucaug gaagcccucuuccucuccca uuucuuccca ucccucaagg 2100 gccuguauug gggcuauguc gguguccucuggugggcgac cauguaccuc ucccaccacu 2160 accucaucga ccucguuggc ggagcuugucugagcgucuu gguguucuac uugugcaugc 2220 cugaagguuu caaggacguu gaucagauacagugggaggc gguggaaggg gauggauacg 2280 agaugauugg cggacccaga acaggcacagguccugagau ugauuuggau gaggaaauua 2340 gaaaguugga agaacaaggc gaagcgcuuuuugagcaggu gauaggggau gaagagucuc 2400 ggaucgagac aaggggagaa ggaaacaguggcgcgggcgg gaaucugagu gggaaugaaa 2460 guggugauag ugcagauagc gguaggggaaagggcaaggg gaaggggaag gggaaacaaa 2520 cugccaaaaa gccaaaggcg aaggagcaaagaucgguuag cuggggagaa acaaagguga 2580 ugggugaagg agcgcaggug gcuucagagaauagcucuua gauaucuguu uuaaaacagu 2640 aguauaccca acaauuuuuu gacgauauacccuaccacua ugaucuuaau aaugcagaau 2700 agaaaaacac cuugccccuc ucgcaggcgucaacacaugg agucuaggca guuaaaucac 2760 acuacaucau caucgccgaa aaaaauaaaccuaguaacuc caacccaacc cuccugguac 2820 aggccuguuc guccaccgac ugauaauucccaucgcccuc gcccuccuca caaguuuacc 2880 cacccgccuu ugcgacuucc aagucaacccggucuccgcc cgacuucuua ucuugcccau 2940 cggguugacg aauucauagg cgaaauuaggguucauaucg ugcaccaagg gguucguuuu 3000 ggugauguaa aaaggaucaa gcuugacggcgauuuucuug ggaggaccaa gaagaggugg 3060 acggggcaaa ggucgagcgg ugggguagauggauucuugg gugaaugcuu gaggggcagu 3120 aaacuagaag aguugagaaa acccaaaggagucagccuuu augucgac 3168

We claim:
 1. A substantially pure IPC synthase protein from a fungalcell other than Saccharomyces cerevisiae.
 2. A substantially pure IPCsynthase protein from a fungal cell comprising an amino acid sequenceselected from the group consisting of: a) SEQ ID NO:2; b) SEQ ID NO:5;c) SEQ ID NO:8; d) SEQ ID NO:11; or e) SEQ ID NO:21.
 3. An isolatednucleic acid encoding a protein of claim
 1. 4. An isolated nucleic acidencoding a protein of claim
 2. 5. An isolated nucleic acid compoundencoding a fungal IPC synthase protein of claim 2, or fragment thereof,wherein said compound has a sequence selected from the group consistingof: (a) SEQ ID NO:1; (b) SEQ ID NO:3; (c) SEQ ID NO:4; (d) SEQ ID NO:6;(e) SEQ ID NO:7; (f) SEQ ID NO:9; (g) SEQ ID NO:10; (h) SEQ ID NO:12;(i) SEQ ID NO:19; (j) SEQ ID NO:20; (k) SEQ ID NO:22; (l) a nucleic acidcompound complementary to (a), (b), (c), (d), (e), (f), (g), (h), (i),(j), or (k); and (m) a fragment of (a), (b), (c), (d), (e), (f), (g),(h.), (i), (j), (k), or (l) that is at least 18 base pairs in length andwhich will selectively hybridize to IPC synthase genomic DNA.
 6. Anisolated nucleic acid compound of claim 5 wherein the sequence of saidcompound is SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ IDNO:19, or SEQ ID NO:20, or a sequence complementary thereof.
 7. Anisolated nucleic acid compound of claim 5 wherein the sequence of saidcompound is SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, or SEQID NO:22, or a sequence complementary thereof.
 8. A vector comprising anisolated nucleic acid compound of claim
 5. 9. A vector, as in claim 8,wherein said isolated nucleic acid compound is SEQ ID NO:1, SEQ ID NO:4,SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:19, or SEQ ID NO:20,operably-linked to a promoter sequence.
 10. A host cell containing avector of claim
 8. 11. A host cell containing a vector of claim
 9. 12. Amethod for constructing a recombinant host cell having the potential toexpress SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, or SEQ IDNO:21, said method comprising introducing into said host cell by anysuitable means a vector of claim
 9. 13. A method for expressing SEQ IDNO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, or SEQ ID NO:21 in therecombinant host cell of claim 10, said method comprising culturing saidrecombinant host cell under conditions suitable for gene expression. 14.A method for identifying inhibitory compounds of fungal IPC synthaseprotein activity, comprising the steps of: a) admixing in a suitablereaction buffer i) a source of IPC synthase protein; ii) a suitablesubstrate; iii) a test inhibitory compound; b) measuring by any suitablemeans an amount of product formed; and c) comparing the amount ofproduct formed at step (b) with a control reaction, said controlreaction comprising steps (a) (i), (a) (ii), and (b) and wherein saidcontrol reaction lacks said test inhibitory compound.
 15. A method, asin claim 14, wherein said synthase protein is selected from the groupconsisting of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQID NO:14, SEQ ID NO:17, and SEQ ID NO:21.
 16. A method, as in claim 14,wherein at step (a)(i) said source comprises membranes from a cell thatexpresses IPC synthase.
 17. A method, as in claim 16, wherein said cellis a recombinant host cell that expresses a vector-borne IPC synthasegene selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, and SEQID NO:20.
 18. An isolated nucleic acid compound for use as a probe orprimer consisting of a nucleic acid molecule wherein the sequence ofsaid nucleic acid is identical with a fragment of or the entirety of SEQID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, or SEQ ID NO:19, or tothe complement thereof.
 19. An isolated nucleic acid compound for use asa probe or primer consisting of a nucleic acid molecule that is at least20 base pairs in length, and wherein the sequence of said nucleic acidis at least 90% identical with a fragment of, or the entirety of, SEQ IDNO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, or SEQ ID NO:19, or to thecomplement thereof.
 20. An isolated nucleic acid compound as in claim 19wherein said nucleic acid hybridizes to said SEQ ID NO:1, SEQ ID NO:4,SEQ ID NO:7, SEQ ID NO:10, or SEQ ID NO:19, or to the complementthereof.